Product Search for novus: Novus Biologicals

AF-401-NA

IL-1 beta/IL-1F2 Antibody [Unconjugated]

(3 Reviews)

(515 Publications)

100 μg

AF-401-NA

Western blot shows lysates of THP‑1 human acute monocytic leukemia cell line untreated (-)ortreated (+) with 200 nM PMA for 24 hoursand 10 μg/mL LPS for 4 hours andRAW 264.7 mouse monocyte/macrophagecell line untreated (-)or treated (+) with10 μg/mL LPS for 24 hours. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Mouse IL‑1 beta /IL‑1F2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-401-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for IL‑1 beta /IL‑1F2 at approximately 35 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

IL‑1 beta /IL‑1F2 was detected in immersion fixed MCF‑7 human breast cancer cell line using Goat Anti-Mouse IL‑1 beta /IL‑1F2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-401-NA) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

IL‑1 beta /IL‑1F2 was detected in perfusion fixed frozen sections of mouse thymus using Goat Anti-Mouse IL‑1 beta /IL‑1F2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-401-NA) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Simple Western lane view shows lysates of RAW 264.7 mouse monocyte/macrophage cell line untreated (-) or treated (+) with 10 µg/mL LPS for 24 hours, loaded at 0.5 mg/mL. A specific band was detected for IL‑1 beta /IL‑1F2 at approximately 40 kDa (as indicated) using 2.5 µg/mL of Goat Anti-Mouse IL‑1 beta /IL‑1F2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-401-NA) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Recombinant MouseIL‑1 beta /IL‑1F2 (Catalog # 401-ML) stimulates proliferation in the the D10.G4.1 mouse helper T cell line in a dose-dependent manner (orange line) as measured by Resazurin (Catalog # AR002). Proliferation elicited by Recombinant Mouse IL‑1 beta /IL‑1F2 (50 pg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse IL-1 beta /IL-1F2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-401-NA). The ND50 is typically ≤0.25 µg/mL.

Quercetin inhibits auto-reactive NLRP3 inflammasome.(A) IL-1 beta concentrations in Nlrp3A350V/A350V and WT BMDM culture supernatants primed with LPS (500 ng/ml) for 90 min and treated with quercetin 1 h after LPS. (B) Supernatants of BMDM were analyzed by anti-ASC, anti-Caspase-1 immunoblotting. Lysates of BMDM were analyzed by anti-IL-1 beta and anti-beta -actin immunoblotting. Data shown are representative of two or more experiments (means ± SD) ***p VC006). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in epithelial cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

GRA7-I-induced inflammasome activation was required for antimicrobial activity in MTB-infected macrophages.(A and B) BMDMs from PKC alpha +/+ and PKC alpha -/- mice (A) and BMDMs were transduced with lentivirus-shRNA-NS or lentivirus-shRNA-ASC for 2 days (B) infected with MTB (MOI = 1) for 4 h and then stimulated with rGRA7 (1, 5, 10 μg/ml) and its mutants for 18 h. IB analysis for IL-1 beta p17, IL-18 p18, or caspase-1 p10 in supernatants (SN), ASC, NLRP3, pro-IL-1 beta , pro-IL-18, or pro-caspase-1 in whole-cell lysates (WCL). Actin was used as a loading control. (C and D) Intracellular survival of MTB was assessed by CFU assay. BMDMs were infected with MTB for 4 h, followed by treatment with rGRA7, and then lysed to determine intracellular bacterial loads. The data are representative of five independent experiments with similar results (A and B). Data shown are the mean ± SD of five experiments (C and D). Significant differences (*P p>0.001,***p<0.001, ns = not significant ) as determined by one way ANOVA with Tukey's post test. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0040722), licensed under a CC-BY license. Not internally tested by R&D Systems.

Effects of IL-1 beta neutralization on the accumulation of MDSCs in lungs.(A) Immunofluorescent photos showed that Gr-1+ (green)/CD11b+ (red) cells indicate MDSCs and increased in lungs of DJ-1 KO mice. Treatment with anti-IL-1 beta neutralizing antibody antagonized the accumulation of MDSCs. DAPI was used to stain nuclei. (B) Bar chart showed the summarized results of MDSCs in lungs of WT and DJ-1 KO mice. Note that the neutralizing antibody significantly reduced the number of MDSCs accumulated in lungs. n = 9 for each group, * p < 0.05 compared with control IgG-treated WT mice, # p<0.05 compared with respective control IgG in WT and DJ-1 KO mice. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0115827), licensed under a CC-BY license. Not internally tested by R&D Systems.

Pro-IL-1 beta degradation is induced by the autophagy inhibitors chloroquine and bafilomycin A, and by the proteasome activator betulinic acid. (A) Chloroquine selectively attenuates IL-1 beta release from cardiac fibroblasts. The cells were incubated with LPS and/or 20 μM chloroquine phosphate as indicated for 18 h, then activated with 3 mM ATP for 60 min. IL-1 beta , TNF, MIP-2, and IL-6 were quantified in the conditioned media (n = 3). (B,C) Cardiac fibroblasts were incubated with LPS for 18 h with or without 20 μM chloroquine phosphate and pro-IL-1 beta quantified with western blot (n = 3). (D) Pro-IL-1 beta mRNA expression was quantified with qPCR (n = 3). (E,F) Cardiac fibroblasts were incubated with LPS for 20 h with or without 10% FCS or 20 μM chloroquine phosphate as indicated. The amount of total ubiquitinated protein was quantified with western blot (n = 4). (G,H) Cardiac fibroblasts were incubated with 10% FCS with or without 10 ng/mL LPS and/or 100 nM bafilomycin A1 for 20 h. Pro-IL-1 beta was quantified with western blot. (I,J) Cardiac fibroblasts were incubated with 10% FCS with or without 10 ng/mL LPS and/or 20 μM betulinic acid as indicated for 20 h. Pro-IL-1 beta was quantified with western blot. All columns are mean with SEM. *p < 0.05, **p < 0.01 (paired student t-test). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31244838), licensed under a CC-BY license. Not internally tested by R&D Systems.

Pro-IL-1 beta maturation is regulated by POH1-mediated deubiquitination. a, b HEK293T cells were transfected with plasmids encoding Flag-caspase-1, Vsv-ASC, His-pro-IL-1 beta (WT) or His-pro-IL-1 beta -K133R with different amounts (125, 250, 500 ng) of plasmid encoding Flag-POH1 and 500 ng of control plasmid, the secretion of a IL-1 beta in the supernatants was quantified by ELISA 36 h after transfection; b the cell lysates were IB with the indicated antibodies. c Mutants of murine pro-IL-1 beta with replacement of various lysine residues. d-f HEK293T cells were transfected with Flag-POH1, V5-caspase-1, V5-ASC, His-pro-IL-1 beta (WT) or its mutants, along d with or e-f without HA-tagged K63-only Ub, then d cells were subjected to IP with an anti-His antibody and IB with the indicated antibodies. e The IL-1 beta levels in supernatants were measured by ELISA; or f the cell lysates were collected and IB with the indicated antibodies. g Proposed function of POH1 as a negative regulator in mediating IL-1 beta processing. POH1, upregulated by TLR3/4 activation, interacts with and deubiquitinates pro-IL-1 beta , resulting in restriction of IL-1 beta cleavage and secretion. Similar results were obtained from three independent experiments. The results represent the mean ± s.d. of three independent sets of experiments. **p < 0.01, ***p < 0.001 (one-way ANOVA with Tukey’s post-hoc test) Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30315153), licensed under a CC-BY license. Not internally tested by R&D Systems.

Serum starvation selectively induces pro-IL-1 beta degradation in cardiac fibroblasts. (A) Cells were incubated with 10 ng/mL LPS for 5 or 20 h with or without 10% FCS and western blot analysis of pro-IL-1 beta and NLRP3 performed. Blots are representative of 3 independent experiments. (B) Protein bands were quantified and the ratios to beta -actin normalized to mean control = 1 (n = 3). (C,D) Cardiac fibroblasts were incubated with or without 10% FCS and primed with LPS for 20 h. Western blot analysis of complex II (mitochondrial mass marker) and NLRP3- inflammasome protein components (NEK7, NLRP3, ASC, pro-caspase-1, and pro-IL-1 beta ) were performed. Bands were quantified and the ratios to beta -actin normalized to mean control = 1 (n = 4, for ASC n = 10). (E,F) Cardiac fibroblasts (120,000 cells per well seeded in 6 well plates) were incubated with 10 ng/mL LPS for 20 h, with or without 10% FCS, then stained with 500 nM MitoTracker Deep Red for 45 min before MitoTracker fluorescence intensity was quantified with flow cytometry analysis (633 nm laser). All cells were analyzed and all gated cells included in the analysis. The presented dot plots (E) show the gate in red and are representative for 9 biological repeats. Median MitoTracker intensity with and without 10% FCS are shown (F). All columns are mean with SEM. *p < 0.05,**p < 0.01 (paired student t-test). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31244838), licensed under a CC-BY license. Not internally tested by R&D Systems.

Differential effects of IL-1 beta neutralization on melanoma lung nodules in WT and DJ-1 KO mice.(A) IL-1 beta antibody (10μg) or control IgG was intravenously injected into mice once. The melanoma cells (6×104) were intravenously injected 30 minutes later. The mice were sacrificed three weeks later. Bar chart showed that the anti-IL-1 beta neutralizing antibody can effectively lower serum IL-1 beta levels as compared with control IgG in both WT and DJ-1 KO mice. Note that serum IL-1 beta levels were higher in DJ-1 KO mice than those in WT mice. (B) Inhibition of IL-1 beta neutralization on lung nodules formation in DJ-1 KO mice. Upper panels: melanoma nodules in lungs of WT (top) and KO (bottom) mice treated with control IgG (left) or neutralizing anti-IL-1 beta antibody (right). For each experimental condition, the gross and microscopic morphologies of tumor nodules are indicated by arrows, and are shown at left and right sites, respectively. Scale bars: 0.5 mm for lung photographs and 0.5 mm for H&E staining. Lower panel: Bar chart showed the summarized results of lung nodule numbers in WT and DJ-1 KO mice. Note that IL-1 beta neutralization enhanced and suppressed melanoma nodules formation in WT and DJ-1 KO mice, respectively. Data are presented as mean ± S.E.M. n = 5 for each group, * p < 0.05 compared with control IgG-treated WT mice, # p<0.05 compared with respective control IgG in WT and DJ-1 KO mice. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0115827), licensed under a CC-BY license. Not internally tested by R&D Systems.

Inflammasome assembly and mature IL-1 beta release in the spinal cord after sciatic nerve axotomy. a Speck-like costaining of NLRP3 and ASC in injured neurons 3 days after the injury (arrows). b Representative WB images of mature IL-1 beta and beta -actin proteins 3 days after unilateral axotomy of sciatic nerve. c Quantification of active IL-1 beta protein expression. N = 3 animals/group. The graph shows values normalized to beta -actin levels and relative to contralateral side. Mean values are shown on bars. Bars represent average ± SEM. **p < 0.01 (ANOVA with Bonferroni post hoc, ax. + vehicle vs. contralateral side). #p < 0.05, ##p < 0.01 (ANOVA with Bonferroni post hoc, compared to ax. + vehicle). Ax. axotomy Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35305649), licensed under a CC-BY license. Not internally tested by R&D Systems.

$GRA7-I activated inflammasomes in an ASC-binding dependent manner.(A) Bacterially purified 6xHis-GRA7-I and its mutants were analyzed by Coomassie blue staining (left) or IB with alpha His (right). (B) BMDMs from PKC alpha +/+ and PKC alpha -/- (left) or BMDMs were transduced with lentivirus-shRNA-NS or lentivirus-shRNA-ASC (MOI = 100) with polybrene (8 μg/mL) (right) for 2 days, the cells were stimulated with rGRA7 (5 μg/ml) and its mutants for the indicated times and culture supernatants were harvested and analyzed for cytokine ELISA for IL-1 beta and IL-18. Data shown are the means ± SD of five experiments. Significant differences (*P < 0.05; ***P < 0.001) compared with PKC alpha +/+ or shRNA-NS. (C) IB analysis for IL-1 beta p17, IL-18 p18, or caspase-1 p10 in supernatants (SN), ASC, NLRP3, pro-IL-1 beta , pro-IL-18, or pro-caspase-1 in whole-cell lysates (WCL). Actin was used as a loading control. (D) IB analysis of lysates of BMDMs as in B and C solubilized with Triton X-100–containing buffer, followed by cross-linkage of insoluble fractions with disuccinimidyl suberate to capture ASC oligomers and analysis of those fractions (I + DSS) and soluble fractions (S) with antibody to ASC. Actin was used as a loading control. (E) Fluorescence confocal images showing formation of speck-like ASC pyroptosomes in BMDMs from PKC alpha +/+ and PKC alpha -/- mice were stimulated with rGRA7 and its mutants for 18 h, fixed, immunostained with antibodies for ASC (Alexa 488). Scale bar, 10 μm. The data are representative of five independent experiments with similar results (A and C-E). Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.ppat.1006126), licensed under a CC-BY license. Not internally tested by R&D Systems.$

The B6 variant of NLRP1B is not cleaved by LF but can form an inflammasome in response to proteolysis.(A) The amino acid sequence of the first 244 residues of NLRP1BB6 was aligned to the homologous sequence of NLRP1B129. The arrow above the alignment indicates the LF-cleavage site in NLRP1B129. Asterisks indicate sites of amino acid identity. (B) For detection of NLRP1B expression, 293T cells were transfected with the indicated amounts of and empty vector (V) or plasmids encoding GFP-HA-NLRP1B129 or GFP-HA-NLRP1BB6 (construct schematics and the predicted molecular weight of each protein is depicted in the upper panel). Cells were treated overnight with anthrax lethal toxin protein (LeTx, 1μg/ml) 24h post-transfection, and then lysates were analyzed by immunoblotting (IB) with the indicated antibodies. For NLRP1 expression and cleavage, CASP1 and IL1B were omitted to prevent cell death and resulting apparent differences of expression. For blots probed with anti-HA (NLRP1B), the lysates were not boiled prior to loading to prevent aggregation and smearing of full-length and FIIND-processed NLRP1B on the immunoblot. To visualize the N-terminal processed form of NLRP1B, the lysates were boiled and resolved on a separate gel. (C) 293T Cells were transfected with the same amount of titrated NLRP1B encoding plasmid and treated as in B, but transfections also included plasmids encoding mouse CASP1 (200ng), IL-1 beta (200ng) and 200ng of empty vector. (D) For detection of NLRP1B expression, 293T cells were transfected for 36h with 250ng of plasmids encoding WT 129 or B6 NLRP1B or mutants engineered to express the TEV-protease site. Each plasmid was co-transfected with either 100ng of empty vector (V) or plasmids encoding TEV-protease (pTEV) or lethal factor protease (pLF), supplemented with 300ng empty vector (pMSCV). Cells were not treated with LeTx and lysates were analyzed by immunoblotting as in (B). (E) Cells were treated as in C, but with 8ng of plasmids encoding NLRP1B, along with 200ng of plasmids encoding mCASP-1 and mIL-1 beta , and 200ng empty vector to normalize plasmid quantities. Lower quantities of NLRP1B were transfected as compared to panel D so to avoid spontaneous NLPR1B activation. For panels B-E, data shown are representative of at least three similar experiments. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.ppat.1006052), licensed under a CC-BY license. Not internally tested by R&D Systems.

SHP inhibits the signal 2 activation of NLRP3 inflammasome.(a,b) LPS-primed BMDMs were treated with fenofibrate (10, 20, 50 μM for 4 h), or (c,d) LPS-primed THP-1 cells were treated with recombinant macrophage-stimulating protein (MSP 10, 50, 100 ng ml−1 for 4 h), and then activated with ATP (5 mM) for 30 min, followed by IB analysis of IL-1 beta p17 or caspase-1 p10 in supernatants (SN), pro-IL-1 beta or pro-caspase-1 in whole-cell lysates (WCL), with actin as a loading control. Data are representative of three independent experiments (a,c). Supernatants were collected and subjected to ELISA for IL-1 beta , IL-18, TNF-alpha and IL-8. Data are the means±s.d. of values from four independent experiments (b,d). *P<0.05; **P<0.01; ***P<0.001, compared with control (two-tailed Student’s t-test). FF, fenofibrate. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms7115), licensed under a CC-BY license. Not internally tested by R&D Systems.

NLRP1A is activated by N-terminal proteolysis.(A) The amino acid sequence of the first 244 residues of NLRP1AB6 was aligned to NLRP1B129 and NLRP1BB6. The arrow above the alignment indicates the LF-cleavage site in NLRP1B129. (B) 293T cells were transfected for 36h with 200ng empty vector (V) or plasmids pcDNA3.1-HA-NLRP1B129 or -NLRP1AB6-MYC (CMV promoter) along with 200ng mCASP1 and 200ng mIL-1 beta expression vectors. Cells were treated overnight with anthrax lethal toxin (LeTx, 1μg/ml) 24h post-transfection, and then lysates were analyzed by immunoblotting (IB) with the indicated antibodies. (C) 293T cells were transfected for 36h with plasmids encoding 400ng GFP-HA-NLRP1B129 or GFP-HA-NLRP1AB6 (under the control of the LTR promoter in pMSCV) or with mutants engineered to contain an N-terminal TEV protease site. Cells were also co-transfected with 100ng empty vector (V) or plasmids encoding TEV-protease (pTEV) or lethal factor protease (pLF), plus 300ng empty pMSCV. (D) To assess IL-1 beta cleavage, cells were transfected as in C, but with 8ng of GFP-HA-NLRP1B and co-transfected with vectors encoding 200ng mCASP1 and 200ng mIL-1 beta . For panels B-D, data shown are representative of at least three similar experiments. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.ppat.1006052), licensed under a CC-BY license. Not internally tested by R&D Systems.

POH1 inhibits inflammasome-induced pro-IL-1 beta processing. a-c BMDMs from Poh1 delta /+ and Poh1 delta / delta mice were primed with LPS for 12–14 h and then stimulated with ATP (0.5 h), nigericin (Nig, 0.5 h), poly(dA:dT) (5 h) or flagellin (5 h) as indicated, a IL-1 beta , b IL-6 and c TNF alpha levels in supernatants were analysed by ELISA. d-f BMDMs lentivirally transduced with control (Vector) or POH1 were stimulated as in a, d IL-1 beta , e IL-6 and f TNF alpha levels in supernatants were analysed by ELISA. g, h BMDMs g derived from Poh1 delta /+ and Poh1 delta / delta mice or h infected by indicated lentiviruses were treated as in a, the cell lysates (CL) and supernatants (SN) were collected and immunoblotted with the indicated antibodies. Similar results were obtained from three independent experiments. The results represent the mean ± s.d. of three independent sets of experiments. **p < 0.01, ***p < 0.001 (two-tailed Student’s t-test) Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30315153), licensed under a CC-BY license. Not internally tested by R&D Systems.

Quercetin inhibits NLRP3 and AIM2 Inflammasomes.LPS (500 ng/ml)-primed BMDM were treated with the indicated flavonoids and concentrations or vehicle (DMSO 0.01%), then stimulated with (A) 5 mM ATP (B) 10 μM Nigericin (C) 130 μg/ml Alum or (D) 400 ng/ml Poly (dA:dT) 30 minutes after treatment. (A–D) IL-1 beta and (E) TNF-alpha concentration in the culture supernatant were measured by ELISA. (B,F,G) LPS-primed BMDM were stimulated with Nigericin treated with quercetin 30 minutes before secondary stimulation. Supernatants and Lysate of BMDM were analyzed by immunoblotting. Cells were stained with FLICA and Caspase-1 activation was determined by FLICA positivity. Data shown are representative of two or more independent experiments (means ± SD) *p < 0.05, **p < 0.01, ***p < 0.001. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28148962), licensed under a CC-BY license. Not internally tested by R&D Systems.

P. gingivalis and its OMVs differentially induce inflammasome signaling and pyroptosis in human macrophages. MDM were infected as before (2 h at MOI of 25:1, see Materials and Methods) with viable P. gingivalis (Pg), heat-killed-Pg (HK-Pg), OMVs, or heat-inactivated-OMVs (HI-OMVs) and (A) the activation of inflammasome components in the lysates was measured after 24 h by Western blot; beta -actin serves as a loading control throughout. Western blot data are representative of at least three independent experiments. (B) Production of IL-1 beta and IL-18 (by ELISA) from MDM was measured in the supernatant at 24 h. Data are mean ± SD from four independent experiments. Cell viability was determined by measuring extracellular LDH release and by 7-AAD exclusion (by flow-cytometry) at 24 h. 7-AAD is a membrane impermeant dye that is excluded from viable cells. The percent of 7-AAD positive (±SD) cells from three independent experiments was calculated relative to the untreated control. MDM treated with H2O2 (1 mM for 60 min) were included as positive control for 7-AAD. A representative histogram is shown. Cells were also treated where indicated with OMVs in the presence of KYT-1 and KYT-36 (10 μM of each). *p < 0.05: n.s, not significant. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28824884), licensed under a CC-BY license. Not internally tested by R&D Systems.

P. gingivalis and its OMVs differentially induce inflammasome signaling and pyroptosis in murine macrophages. BMM were infected as before (2 h at MOI of 25:1, see Materials and Methods) with viable P. gingivalis (Pg), heat-killed-Pg (HK-Pg), OMVs, or heat-inactivated-OMVs (HI-OMVs) and (A) the activation of inflammasome components in the lysates [or supernatants (sup) where indicated] measured after 24 h by Western blot; beta -actin serves as a loading control throughout. Western blot data are representative of at least three independent experiments. (B) Production of IL-1 beta and IL-18 (by ELISA) from BMM was measured in the supernatant at 24 h. Data are mean ± SD from four independent experiments. Cell viability was determined by measuring extracellular LDH release and by 7-AAD exclusion (by flow-cytometry) at 24 h. 7-AAD is a membrane impermeant dye that is excluded from viable cells. The percent of 7-AAD positive (±SD) cells from three independent experiments was calculated relative to the untreated control. BMM treated with H2O2 (1 mM for 60 min) were included as positive control for 7-AAD. A representative histogram is shown. Cells were also treated where indicated with OMVs in the presence of KYT-1 and KYT-36 (10 μM of each). *p < 0.05. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28824884), licensed under a CC-BY license. Not internally tested by R&D Systems.

Caspase-1 but not gasdermin-D processing depends on ASC oligomerization.(a) Western blot analysis for cleaved caspase-1 p20, IL-1 beta p17, and HMGB-1 in cell supernatants (SN) and pro-caspase-1, pro-IL-1 beta and HMGB-1 in cell lysates (lys) of LPS-primed immortalized Asc−/− BMDMs or Asc−/− BMDMs expressing ASCFL, ASCY59A or ASCE80R 3 h after poly(dA:dT) transfection (1 μg ml−1). (b) Release of LDH from LPS-primed immortalized Asc−/− BMDMs or Asc−/− BMDMs expressing ASCFL, ASCY59A or ASCE80R, or derived Casp1 knockouts 3 h after poly(dA:dT) transfection (1 μg ml−1). (c) Western blot analysis for processing of full-length gasdermin-D (GSDMDFL) into the active N-terminal fragment (GSDMDN-term) in combined lysates and supernatants (lys + SN) of LPS-primed immortalized Asc−/− BMDMs expressing ASCFL, ASCY59A or ASCE80R, or derived Casp1 knockouts 3 h after poly(dA:dT) transfection (1 μg ml−1). Arrowhead, gasdermin-DNterm p30; *a cross-reacting band. (d) Release of LDH from LPS-primed primary C57BL/6 WT (WT), Casp1−/−/Casp11−/− or Asc−/− BMDMs infected with S. Typhimurium (multiplicity of infection (MOI)=10, 1 h). (e) Western blot analysis for processing of full-length gasdermin-D (GSDMDFL) into the active N-terminal fragment (GSDMDN-term) in combined lysates and supernatants (lys+SN) of LPS-primed primary C57BL/6 WT (WT), Casp1−/−/Casp11−/− or Asc−/− BMDMs infected with S. Typhimurium (MOI=10, 1 h) or left uninfected. Arrowhead, gasdermin-DNterm p30; *a cross-reacting band; ** a S. Typhimurium-specific cross-reactive band. See also Supplementary Figs 8 and 9. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms11929), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

, , , ,

Applications: , , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Biotin	BAF401

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF-401-SP

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NB400-105

ABCA1 Antibody - BSA Free

(7 Reviews)

(398 Publications)

0.1 ml

NB400-105

Formalin-fixed paraffin-embedded tissue sections of human prostate cancer were probed for ABCA1 mRNA (ACD RNAScope Probe, catalog # 432291; Fast Red chromogen, ACD catalog # 322360). Adjacent tissue section was processed for immunohistochemistry using rabbit polyclonal (Novus Biologicals catalog # NB400-105) at 1.5ug/mL with overnight incubation at 4 degrees Celsius followed by incubation with anti-rabbit IgG VisUCyte HRP Polymer Antibody (Catalog # VC003) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to glandular cells.

<b>Genetic Strategies Validation and Orthogonal Strategies Validation.</b>HSKMCs were transfected with scrambled siRNA (scr siRNA) or ABCA1 siRNA. The cells were lysed and ABCA1 (A) protein levels were quantified by immunoblotting. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-018-38014-3) licensed under under a CC-BY license.

<b>Biological Strategies Validation. </b>Analysis of ABCA1 in total cell lysates of RAW264.9 cells treated with vehicle (-) or 9-cisretinoic acid and 22Rhydroxycholesterol (+). Samples used for this testing were 40 ug of total cell post-nuclear lysate from each group.

<b>Biological Strategies Validation. </b>HepG2 cells were grown to 60% confluency, serum starved for 24 hours, and then treated with 1uM TO9 for 24 hours prior to being fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-ABCA1 at 5.0ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Untreated HepG2 cells were grown to 60% confluency, and serum starved for 24 hours prior to being fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-ABCA1 at 5.0ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

ABCA-1 FITC conjugated antibody of human adipose tissue macrophage subsets by flow cytometry. Image from verified customer review.

Association between expression of ABCA1 and survival in ovarian cancer patients. Expression of ABCA1 in 55 ovarian cancer patient samples was determined by IHC in tissue microarray. Representative image of ovarian cancer showing high (left panel) and low (right panel) ABCA1 expression on the cell membrane or cytoplasm (x400). Image collected and cropped by CiteAb from the following publication (//www.clinicalepigeneticsjournal.com/content/7/1/1), licensed under a CC-BY license.

ABCA1 in human primary fibroblasts in culture. 10 ug of total protein. 7.5% TGX gel. Blocking 5% milk in PBST 1h RT. 1:1000 primary ab in BSA 3% PBST O/N at 4C. Secondary 1:5000 HRP 1h RT. Arrow shows around 250 kDa. WB image submitted by a verified customer review.

ABCA1 antibody was tested at 1: 400 in HeLa cells using an Alexa Fluor 488 secondary (shown in purple). M1 is defined by unstained cells.

<b>Biological Strategies Validation. </b>Detection of ABCA1 in mouse peritoneal macrophages using NB 400-105 (Lot L). ECL exposure, 1 min. Lane 4: T09 uninduced lysate Lane 5: T09 induced lysate.

Prostates of LXR mutant mice accumulate cholesterol esters through inappropriate LXR target genes regulation. Total protein lysates of WT and LXR null mice under normal or high cholesterol diet were analyzed by western blotting with antibodies against ABCA1, LDLR and ACTIN as a loading control (left panel), quantification of ABCA1 and LDLR protein accumulation levels (right panel). * p<0.05, *** p<0.001 in Student's t test. Error bars represent the +/- mean SEM. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pgen.1003483), licensed under a CC-BY license.

miR-33b reduces cellular cholesterol efflux and serum HDL-C levels. Western blotting for ABCA1 (using NB400-105) and ABCG1 proteins in peritoneal macrophages from WT and KI+/+ mice. Representative images are shown. beta-actin was used as the loading control. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep05312), licensed under a CC-BY license.

miR-33b regulates ABCA1 and SREBP-1. Analysis for ABCA1, SREBP-1, CPT1a, and AMPK alpha protein levels in the livers of WT, KI+/-, and KI+/+ mice. Representative images are shown. TF2B and beta-actin were used as loading controls. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep05312), licensed under a CC-BY license.

Detection of ABCA1 in human prostate epithelium showing luminal and membrane staining.

Prostates of LXR mutant mice accumulate cholesterol esters through inappropriate LXR target genes regulation.(A) Plasma concentrations of cholesterol were determined (N = 9/13 per group) after 5 weeks dietary conditional exposure in each genotype. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23675307), licensed under a CC-BY licence.

Gas6 treatment enhances expression of LXRalpha and LXRbeta and their target genes in BMDM.Mouse BMDM were stimulated with 400 ng/ml Gas6, 1 uM T0901317, 10 ng/ml interferon (IFN)-gamma, 10 ng/ml IL-4, or 100 ng/ml LPS for 4 h (a,b) or 400 ng/ml Gas6 for the indicated times (c-g). (a,b,e) The amounts of the LXRalpha, LXRbeta, ABCA1, ABCG1, ApoE, AIM, Arg2, VEGF, YM1, and Arg1 mRNAs were analyzed by real-time PCR and normalized to that of Hprt mRNA. (c,d,f,g) The relative abundances of LXRalpha, LXRbeta, ABCA1, ABCG1, ApoE, Aim, Arg2, and VEGF proteins were determined by Western blotting analysis. The relative densitometric intensity was determined for each band and normalized to beta-actin. Data in all bar graphs are means +/- SEM of three independent experiments. *P < 0.05 compared with control. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27406916), licensed under a CC-BY licence.

Western Blotting for ABCA1, as detailed in Supplementary Protocol #5 (Additional file 1). Caco-2 cells cultivated on filter inserts were treated for 48 h with bexarotene (RXR agonist, 5 uM), T0901317 (LXR agonist, 10 uM), GW3965 (LXR agonist, 10 uM) or the vehicle control (DMSO 0.1%). Protein expression was then determined by western blot analysis. Results were normalized for interexperimental variance and then set in relation to the vehicle control. Bar graphs represent mean +/- SD from three independent experiments. A representative blot is shown - the bands stem from the same membrane and the vertical line indicates excised irrelevant bands. ns not significant, *P < 0.05, **P < 0.01 versus solvent vehicle (determined by paired t-test) Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32308567), licensed under a CC-BY licence.

Anti-ApoA-1 IgGs upregulate ABCA1. ABCA1 was increased after 16 and 24 h of anti-ApoA-1 IgG exposure to HMDM at the mRNA level as revealed by RT-PCR (a) as well as at the protein level after 24 h anti-apoA-1 IgG stimulation, as revealed by Western blot analysis (b). In panel a, data are expressed as fold change expression of the mean +/- SD of ABCA1 calculated by deltadeltaCT method of six independent experiments (n = 6) and values were normalized to untreated condition. p-values were calculated using the Student's t-test: * p < 0.05, ** p < 0.01, **** p < 0.0001. (b) One of four representative Western blots is shown. Data are the mean +/- SD of band intensity volume/actin intensity volume of four independent experiments (n = 4). p-values were calculated using the Student's t-test: * p = 0.02, ** p = 0.005. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31766415), licensed under a CC-BY licence.

ApoA-I increases glucose uptake in HSKMCs in an ABCA1- and SR-B1-dependent manner. HSKMCs were transfected with scrambled siRNA (scr siRNA), ABCA1 siRNA or SR-B1 siRNA. The cells were lysed and ABCA1 (A) and SR-B1 (B) protein levels were quantified by immunoblotting. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30718702), licensed under a CC-BY licence.

Functional miR-33 responsive elements in the 3'UTR of ATP8B1 and ABCB11. G). Relative protein levels in HuH7 cells transduced with empty or miR-33 adenovirus. Some cells were incubated for 16 h in the presence of FXR:RXR agonists (2 umol/L GW4064 : 1 umol/L 9-cis-retinoic acid) to induce ABCB11. Asterisk indicates a non-specific band. Data shown as mean +/- SD; **p < 0.01. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/22767443), licensed under a CC-BY licence.

LXRs control expression of genes involved in cholesterol homeostasis and fatty acid synthesis in MPECs. (D) Western blot analysis was performed on WT (+/+) or Lxralphabeta-/- MPECs using Srebp1c, Abca1 and beta-Actin antibodies. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23554947), licensed under a CC-BY licence.

Suppressive effects of Ucn1 on foam cell formation and related protein expression in human monocyte-derived macrophages.Human monocytes were incubated for 7 days with RPMI-1640 supplemented with 10% human serum and the indicated concentrations of Ucn1, followed by a 19 h-incubation with 50 ug/ml oxLDL in the presence of 0.1 mmol/l [3H]oleate. Intracellular CE accumulation was determined from the radioactivity of cholesterol-[3H]oleate. Otherwise, before the addition of oxLDL, cells were harvested and subjected to immunoblotting analyses for CD36, ACAT1, or ABCA1. beta-Actin served as a loading control. Data are expressed as means +/- SEM from 4-6 independent experiments with monocytes from 4-6 different donors. Baseline (1 fold) = 8.4+/-2.2 nmol/mg cell protein. *P<0.05, +P<0.001 vs. 0 nmol/l of Ucn1. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0110866), licensed under a CC-BY licence.

Alternatively activated macrophages (M2) exhibit increased clearance of OxLDL in vitro C). Increased ABCA1 and ABCG1 expression in M2-derived foam cells. Shown is a representative Western blot for the presence of ABCA1, ABCG1 and beta-actin in lysates of cells that were treated as indicated. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23027612), licensed under a CC-BY licence.

$Impairment of mitophagy upon Antimycin A and Oligomycin exposure.a Cells were treated with Antimycin A (1 μM) and Oligomycin (1 μM) for 1, 3, and 6 and mitochondrion/cytosol fractionation was performed to assay the accumulation of LC3-II, p62 and PINK1 within the mitochondrial fraction. VDAC1 was used as a loading control for mitochondrial fractions. b, c Cells were treated with Antimycin A (1 μM) and Oligomycin (1 μM) for 1, 3, 6, and 24 h and total fibroblasts extracts were analyzed for PINK1, p62, COXIV and VDAC1. Each quantitative data was normalized with ACTIN (n ≥ 3). Statistical analysis was performed using two-way ANOVA with Tukey’s multiple comparisons test. (*p < 0.05; **p < 0.01; ***p < 0.001). Immunoblots reported are from one representative experiment Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31332166), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

GPR50 competes with FKBP12 for the binding to T beta RI. a (Left) HEK293T cells were transfected with Flag-T beta RI alone or cotransfected with HA-T beta RII (with TGF beta ; 2 ng/mL; 1 h), myc-FKBP12 and either GPR50 delta 4 or GPR50wt. Lysates were precipitated for FKBP12 using anti-myc antibody and blotted with an anti-Flag to reveal complex formation. Expression of myc-FKBP12, Flag-T beta RI, HA-T beta RII and GPR50 was determined in total lysates. (Right) Densitometric analysis of 3 independent experiments (Mean ± s.e.m., n = 3 independent experiments, **p < 0.01, one-way ANOVA with Dunnett’s post hoc test). b, c Competition between FKBP12 and GPR50 for T beta RI binding was checked by precipitating FKBP12 from total brain and lung lysates of wt and GPR50ko mice and revealed with anti-T beta RI. Total lysate was addressed for expression of FKBP12, T beta RI, and GPR50 with corresponding antibodies. d To address the competition of FKBP12 and GPR50 for T beta RI binding, BRET measurements were performed with HEK293T cells expressing fixed amounts of T beta RI-Rluc8 and GPR50 delta 4-YFP or GPR50wt-YFP or T beta RII-YFP and stimulated with TGF beta (0.6 nM, 30 min, 37 °C). Immunoblot on the top shows FBP12 expression when transfected either with empty (Mock) or FKBP12 in different conditions. e HEK293T cells expressing the indicated proteins were starved and treated for 1 h with 2 ng/ml of TGF beta or 100 ng/ml of FK506. Total lysates were immunoblotted for Smad3 phosphorylation and total expression of myc-Smad3, GPR50 and myc-FKBP12 with suitable antibodies. f Alignment of FKBP12 and GPR50 sequences revealed similarities between the C-terminal “ATGHP” motif in FKBP12 and four repetitive motifs in GPR50 (upper top panel). One motif of GPR50 is located close to the delta 4 deletion of GPR50 delta 4 (lower bottom panel). Structural data with permission adapted from Huse et al.29 highlight the implication of the HP loop (red) in binding to T beta RI (lower panel). g HEK293T cells were transfected with indicated plasmids and as in a. Lysates were precipitated for FKBP12 using an anti-myc antibody and blotted with an anti-T beta RI to reveal complex formation. Expression of myc-FKBP12, HA-T beta RI, and GPR50 was determined in total lysates. Representative results are shown for b, c, e, and g. Similar results were obtained in at least two additional experiments. See also Supplementary Fig. 3 Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/s41467-018-03609-x), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Involvement of CBP in BNIP3 expression regulated by HIF-1 alpha and FOXO3 under hypoxia. (A) UCB-hMSCs were incubated with hypoxia condition for 24 h. Co-immunoprecipitation of HIF-1 alpha and FOXO3 with IgG and CBP were shown in left panel. IgG was used as a negative control. The total protein expressions of HIF-1 alpha , FOXO3, CBP and beta -actin in lysate were shown in right panel. n = 3. (B) CBP (20 μM) was pretreated to UCB-hMSCs, and cells were incubated with hypoxia for 24 h. The BNIP3 mRNA expression level was analyzed by qPCR. n = 6. (C) BNIP3 and beta -actin protein expressions were analyzed by western blot. Data represent mean ± S.E. n = 4. (D, E) Sample DNA was immuno-precipitated with RNA polymerase, IgG, HIF-1 alpha and FOXO3 specific antibodies. CHIP (top panel) and lysate (bottom panel) samples were amplified with the primers of GAPDH and BNIP3 promoters. Quantitative CHIP data was analyzed by qPCR, and shown in the right panel. n = 4. Western blot data were normalized by beta -actin, and qPCR data were normalized by ACTB mRNA expression level. Quantitative data are presented as a mean ± S.E.M. All blot images are representative. *p < 0.05 versus control, #p < 0.05 versus hypoxia. Image collected and cropped by CiteAb from the following open publication (//linkinghub.elsevier.com/retrieve/pii/S2213231717303804), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

DNA methylation and expression level of the PHD3 gene in HCT116 and DLD-1 CRC cells. A. HCT116 and DLD-1 cells were cultured under normoxic or hypoxic (1% O2) conditions for 48 hrs. Cells were then used for DNA isolation followed by bisulfite modification. Methylation percentage of three DNA fragments within the PHD3 CpG island (Additional file 1, Additional file 2) in HCT116 and DLD-1 cells under hypoxic and normoxic conditions was determined by Real Time PCR amplification of bisulfite treated standard and cell line DNA, followed by comparison of their HRM profiles. B. Cells were cultured in DMEM either in hypoxic (1%O2) or normoxic conditions for 48 hrs. After incubation, the cells were used for total RNA isolation and reverse transcription. The PHD3 cDNA levels were determined by RQ-PCR relative quantification analysis. RQ-PCR results were standardized by the geometric mean of PBGD and hMRPL19 cDNA levels. PHD3 cDNA levels are expressed as a multiplicity of these cDNA copies in the cell line’s calibrator. C. Cells were cultured in DMEM either in hypoxic (1%O2) (H) or normoxic (N) conditions for 48 hrs. Cells were then used for protein isolation. Proteins were separated by 10% SDS-PAGE, and transferred to a membrane that was then immunoblotted with Rp anti - PHD3 Ab and incubated with goat anti-rabbit HRP-conjugated Ab. The membrane was then stripped and reblotted with Rp anti-GAPDH Ab, followed by incubation with goat anti-rabbit HRP-conjugated Ab. The band densitometry readings were normalized to GAPDH loading control. The ratio of PHD3 to GAPDH for DLD-1 in normoxic conditions was assumed to be 1. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/24195777), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

The role of HIF-1 in purinosome formation.a, quantifying the number of purinosome-containing cells in normoxia or hypoxia (24 h) in purine-rich medium and normoxia in purine-depleted medium (purine -ve), cells in hypoxia transfected with siRNA to HIF-1 alpha (+ siRNA), and cells in purine-rich medium supplemented with DFX. Data shown are n = 3, mean ± S.E., total number of cells counted are shown in parentheses. b, time course of purinosome formation in hypoxia shows the number of purinosome-containing cells steadily increases after 3 h in hypoxia. Re-oxygenation of the samples after hypoxic incubation for 10 h reverts the number purinosome-containing cells back to normoxic levels. Data shown is n = 3, mean ± S.E., total number of cells counted are shown in parentheses. c, time course of HIF-1 alpha stabilization in hypoxia shows maximum HIF-1 alpha protein expression levels at 3 h in hypoxia, after which the HIF-1 alpha expression decreases. The positions of molecular markers are shown for each blot; uncropped blots with overlaid markers are deposited in the raw data files. d, the effect of hypoxia on the transcription of purine biosynthesis enzymes measured by qPCR. Vascular endothelial growth factor (VEGF) and HIF-1 alpha are controls. Data shown are n = 5, mean ± S.E. e, the effect of hypoxia on the protein expression levels of the purine biosynthetic enzymes. HIF-1 alpha is stabilized in hypoxia as expected, and no significant increase in the purine enzymes was detected between normoxic (21% oxygen) and hypoxic (1% oxygen) growth conditions. The positions of molecular markers surrounding each band of interest are shown for each blot; uncropped blots with overlaid markers are deposited in the raw data files. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32439803), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

FIH exits the nucleus via a Leptomycin B-sensitive exportin1-dependent pathway. (A) Immunofluorescence staining of FIH (green) and HIF1 alpha (red) in MCF7 cells after the indicated hypoxia (0.5% O2) and re-oxygenation treatments. TO-PRO-3 (blue) was used to stain nuclei. (B) Total cell lysates from U2OS cells were immunoprecipitated with an anti-exportin1 antibody or control IgG. FIH, exportin1 and beta -tubulin levels are indicated. (C) Immunofluorescence staining of FIH (green) in FIH-null mouse embryonic fibroblasts (MEFs) transfected with HA-FIH 1–349 or HA-FIH delta NES followed by normoxia, hypoxia (1% O2, 3 h) or 3 h of hypoxia followed by re-oxygenation for 1 h. TO-PRO-3 (blue) was used to stain nuclei. Arrows indicate nuclear localization of signal. (D) Total cell lysates from U2OS cells transfected with control vector, HA-FIH 1–349 or HA-FIH delta NES were immunoprecipitated with an anti-exportin 1 antibody. HA-FIH, exportin1 and beta -tubulin levels are indicated. FL, full length; IgGL, IgG light chain. Scale bars: 20 µm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30333145), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Munc13-4 KD strongly impairs exosome release. (A) SDS-PAGE Western blot of indicated proteins in MDA-MB-231 cells after stable expression of shRNA for Munc13-4 or Rab27a or a scrambled control (Ctrl). (B) Culture medium from MDA-MB-231 cells either untreated or stimulated with 1.25 µM ionomycin for 30 min was centrifuged at 1,000 g to remove cellular debris and 10,000 g to remove large extracellular vesicles. (C) The resulting 10,000-g supernatant was filtered onto a nitrocellulose membrane and analyzed for CD63, CD9, ALIX, and GM130 content by antibody blotting. (D) Quantification of CD63, CD9, and ALIX blots in C are shown as exosome release as a percentage of total cellular material with mean values ± standard error (SE) for n ≥ 3. *, P < 0.05 for comparison with corresponding control samples. (E) Panc-1 or A549 cells were left untreated or were treated with TGF beta -1 for 24 h. Indicated proteins were detected by SDS-PAGE Western blot. (F) Panc-1 cells were left untreated or were treated with TGF beta -1 for 24 h, and Munc13-4 levels were determined by immunofluorescence. TGF beta -1–treated cells exhibited a mesenchymal morphology. Bars, 5 μm. (G) A549 cells stably expressing control shRNA (Ctrl) or Munc13-4 shRNA were left untreated (Untr) or were treated with TGF beta -1 for 24 h, and SDS-PAGE Western blotting for indicated proteins was conducted. (H) Culture media supernatants (as in B) from A549 cells that were either untreated or were stimulated with 1.25 µM ionomycin for 30 min were filtered onto nitrocellulose membrane and analyzed for CD63 and GM130. (I) Quantification of CD63+ exosome release shown as a percentage of total cellular material with mean values ± SE for n = 5. *, P < 0.05; **, P < 0.01 for comparison with corresponding control samples. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29930202), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Elevated Spy1 levels prevent contact inhibition. (A) Representative views of focus formation assays in NIH3T3 cells transfected with Myc-Spy1-WT, Myc-Spy1-TST, positive control Ras-V12 or empty vector PCS3 control. n = 3 (B) Quantification of the number of colonies. Error bars reflect SE between triplicate experiments. t test was performed;** P ≤ 0.01. n = 3 (right panel) Western blot analysis of one representative lysate from A-B. Image collected and cropped by CiteAb from the following open publication (//bmccancer.biomedcentral.com/articles/10.1186/1471-2407-12-45), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

SOD2 mRNA and protein expression. RT-PCR: (A) SOD2 mRNA expression is higher in adults, and highest in the adult parenchyma. (B) No exposure effects on SOD2 mRNA were observed in neonates. (C) Adult SOD2 mRNA was decreased in PFP48. Data are presented as mean+SEM (n=5-7 rats/group, in each compartment), * significantly different compared to neonates in the same compartment, † significantly different compared to airways in the same age, ‡ significantly different compared to FA in the same compartment. Western blotting: (D) Scan of representative SOD2 and actin blots. (E) Neonatal whole lung SOD2 protein expression was unchanged with exposure, and (F) adult whole lung SOD2 protein trended upwards at PFP2, but was statistically insignificant. (G-J) Immunohistochemical localization of SOD2 in lung (n=6 rats/group). SOD2 protein was more abundant in adults compared to neonates, but no exposure specific differences were observed. Scale bar is 50 μm. Image collected and cropped by CiteAb from the following open publication (//particleandfibretoxicology.biomedcentral.com/articles/10.1186/1743-8977-10-34), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Cells of human meninges co-express LLEC markers. a–c DAB-IHC with single antibodies detects VEGFR3 (a), LYVE1 (b), and MRC1 (c) in the meninges of human post mortem brain showing no signs of neuropathology. These images are taken from a 38 year old male (sample P17/07, Table 1), and confirmed in n = 2 additional samples. P parenchyma. Scale = 150 µm (a); 40 µm (b); and 20 µm (c). d–f DAB-IHC with single antibodies detects VEGFR3 (b), LYVE1 (c), and MRC1 (d) in elderly human meninges (age: 89–92) with evidence of neuropathology and confirmed in n = 3 brains (Table 1). P, parenchyma. Scale = 20 µm. g–p IHC with fluorescent antibodies detects human meningeal cells that co-express MRC1 (h, m, yellow), LYVE1 (i, n, white), and VEGFR3 (j, o, green). Nuclei/RNA are labelled with DAPI (g, l, blue) and images are merged in (k, p). Scale = 10 µm Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31696318), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Inhibition of KDM expression in vivo.(A) Ears from C57Bl/6 mice were injected intradermally with 250 ng of IL-1 alpha , IL-17A, IL-22, OSM and TNF alpha (M5) or with Phosphate Buffered Saline (PBS). At 24 h quantitative RT-PCR analysis was carried out on total RNA and expression levels for cytokeratin 1 (CK1), cytokeratin 10 (CK10), loricrin (LOR), filaggrin (FLG), desmoglein 1 (DSG1), desmocollin 1 (DSC1) and involucrin (IVL) were normalized using GAPDH housekeeping gene and expressed as the fold decrease under non injected skin. Data are represented as mean and SEM of 3 independent experiments. *p<0.05, **p<0.01, ***p<0.001. (B) On day 2, the ears were collected for staining with Hematoxylin and Eosin (HE) and immunodetection of cytokeratin 10, loricrin, filaggrin, cytokeratin 6 and Ki-67. Scale bar 100 µm. Results are from one experiment representative of three. (C) Skin biopsies from normal control skin (Cont) or lesional psoriatic skin (Pso) were collected. Skin sections were stained with Hematoxylin and immunodetection of cytokeratin 10, loricrin, filaggrin, involucrin and S100A7 was performed. Scale bar 100 µm. Results are from one experiment representative of three. Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.pone.0101937), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

MicroRNA-214-3p (miR-214) mimic in rat vascular smooth muscle cells (A7r5) suppressed angiogenesis and proliferation but promoted senescence. a Schematic of the protocol for transfection of A7r5 VSMCs with a miRNA mimic control (mimic NC) or a miR-214 mimic and processed at the indicated times. b The effects of miR-214 mimic on VSMC cell proliferation were tested by CCK-8 assay. c Angiogenetic mRNA expressions of NOS3, VEGFA, CXCL12 and CXCR4 compared in cells transfected with mimic NC or miR-214 mimic (n = 3). d Senescence-associated mRNA expressions of TERT, TERF1 and TERF2 compared in cells transfected with mimic NC or miR-214 mimic (n = 3). e,g Representative western blots depicting TERF1, TERF2, p16INK4, p21CIP1, pRB, and quaking expression in mimic NC or miR-214 mimic transfected cells. f,h Normalized expressions of TERF1, TERF2, p16INK4, p21CIP1, pRB, and quaking (n = 3). i Senescence-associated beta -galactosidase staining demonstrating senescence in mimic NC or miR-214 mimic transfected cells. j Bar graphs show quantification of relative of SA-beta -gal positive cells (n = 3). Data are presented as the means ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001 (Two-tailed Student’s t-test) Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32410577), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Intraperitoneally Administered Compound A Effectively Suppresses Triple-Negative Breast and Blood Cancer Xenograft Tumor Growth. Immunocompromised mice were subcutaneously injected with cancerous cell lines and tumors were allowed to establish. Treatments occurred every other day and the studied compound or the equivalent vehicle control administered intraperitoneally for five weeks. Tumor volume and mass were measured two times per week. (a) MV-4-11 xenograft study was administered with approximately 5.0 mg/kg of Compound A and 5.0 mg/kg of curcumin. Results are representative for the Control (n = 5), Compound A (n = 5), and curcumin (n = 4) treated group, each with two tumors per mouse. (b) MDA-MB-231 xenograft study was administered with approximately 7.5 mg/kg of Compound A. Results are representative for the Control (n = 3) and Compound A (n = 4), each with one tumor per mouse. Scale bar is 1 centimeter. (c) Immunohistochemistry analysis of sectioned tumor tissues from the MDA-MB-231 study. Each section was subjected to the specified antibody followed by a biotinylated secondary antibody. Detection was done using a DAB Peroxidase HRP Substrate Kit (brown) followed by Hematoxylin counterstaining (purple). Images were obtained using inverted bright field microscopy. Sectioning results are representative of three individual tumors. Scale bar is 50 microns. Statistical analysis using One-Way ANOVA. *p < 0.05 vs tumor volume of the control. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28439094), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Mechanical tensile strain of 30% induces an elongation of paranodal junction and alteration in MAPK signaling in organotypic cerebellar slice culture. a Double immunostaining of organotypic cerebellar slices for CASPR in red and MAG in green. The response was evaluated at time 0 h post-stretch. Altered paranodal junctions are highlighted within white circles in high-magnification images. Scale bar, 50 μm; inset scale bar, 5 μm. b Sketch of CASPR and MAG labeling in the paranodal junction. Quantitative analysis is characterized by three length measurements: c nodal length called (A); d paranodal length called (B); and e (B-A) corresponding to the total length of the CASPR labeling. f–h Proteins were extracted following strain of 30% from control (Ctrl) and stretched (Stretch) slices and western blot was performed for the myelin protein MBP (f), ERK1/2 (g), and p38 (h). The ratio P-ERK on total ERK and the ratio P-p38 on total p38 are presented. beta -Actin was used as a loading control. Results represent the mean ± SEM (n = 3). ***p < 0.001. CASPR contactin-associated protein, MAG myelin-associated glycoprotein Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30298339), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Gomisin N (GN) represses the expression of JMJD2B, PPAR gamma 2, and PPAR gamma 2 steatosis target genes in HepG2 cells. HepG2 cells were treated with GN (100 μM) for 12 h. (A) JMJD2B expression was assessed by qPCR and western blotting. The full-length western blots corresponding to truncated blots are given in Supplementary Figure S4. (B) PPAR gamma 2 expression was measured by qPCR. (C,D) The expression of PPAR gamma 2 steatosis target genes was assessed by qPCR. Data represent means ± SEM of three independent experiments performed in triplicate. *p < 0.05 vs. no treatment. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30214048), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

TCAB1 knockdown inhibits tumor growth in vivo. a. shTCAB1-C and shScra cells were injected subcutaneously into the left and right neck of the nude mice respectively. Measured the tumors size from the 11th day using vernier caliper and calculated the volume of tumors by formula l*w*h/2 (mm3). Finally, got out the xenografts carefully after mercifully killed the mice. b. Measured the final volume and weighted the final weight of the removed tumors. c. Performed IHC against TCAB1 using mice xenografts sections. The smaller tumors expressed less TCAB1 compared to shScra cells. Statistical analysis of the IHC data used Aperio ImageScope software and all of the data were determined by Student’s t test (*P < 0.05, **P < 0.01, ***P < 0.005). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/25070141), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

MiR-375-3p negatively regulates Derlin-1 and blocks EMT in BFTC909 cells. (A) Western blot revealed the restoration of Derlin-1, MMP-2, Snail, and ZEB1 after co-transfection of miR-375-3p mimics and CMV-Derlin-1 compared with cells transfected with miR-375-3p alone in BFTC909 cells with alpha -tubulin as a reference (B) Quantification of the protein levels of Derlin-1, occludin, MMP-2, Snail, and ZEB1 from (A) (N = 3). (C) miR-375-3p suppressed BFTC909 cell migration ability but restored by Derlin-1 overexpression (N = 3). (D) miR-375-3p repressed invasion of BFTC909 cells but restored by Derlin-1 overexpression (N = 3). Data were represented as mean ± SD; * p < 0.05, ** p < 0.01. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35205628), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

JMJD2B expression increases in hepatic steatotic cell and animal models. (A) HepG2 cells were incubated with a mixture of palmitic acid (PA) and oleic acid (OA) (1:2 ratio) at 800 μM concentrations for 24 h, and intracellular triglyceride (TG) levels were analyzed by a TG assay kit. JMJD2B mRNA and protein levels were examined by qPCR and western blotting, respectively. Data represent means ± SEM of three independent experiments performed in triplicate. *p < 0.05 vs. no treatment. The full-length western blots corresponding to truncated blots are presented in Supplementary Figure S1A. (B) HepG2 cells were treated with T0901317 (10 μM) for 24 h, and intracellular triglyceride (TG) levels were measured by a TG assay kit. JMJD2B mRNA and protein levels were examined by qPCR and western blotting, respectively. Data represent means ± SEM of three independent experiments performed in triplicate. *p < 0.05 vs. no treatment. The full-length western blots are presented in Supplementary Figure S1B. (C) Total RNAs were isolated from the livers of HFD-induced obese mice. The JMJD2B mRNA levels were assessed by qPCR. Data represent means ± SEM of 5 mice. *p < 0.05 vs. ND mice. ND: normal diet. HFD: high fat diet. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30214048), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

TLR4 expression in pVW-MSCs cultured with or without LPS (10 μg/ml) for 1 and 4 h and after additional 24 h of recovery after LPS removal (4hR). a: flow cytometry analysis were performed in not fixed and not permeabilized cells for TLR4 surface expression determination (TLR4: Surface) and in fixed and permeabilized cells to measure the overall TLR4 amount (TLR4: Total). Red histograms: stained cells; blue histograms: control cells. b: representative Western Blot of TLR4 and housekeeping beta -tubulin and relative quantification were presented. c: representative images of TLR4 immunostaining of pVW-MSCs cultured with or without LPS (10 μg/ml) for 4 h. pVW-MSCs nuclei were stained with Hoechst 33258 (blue). Scale bar = 10 μm. Data shown represent the mean ± SD of three biological replicates, each experiment is repeated three times. Data were analysed using one-way ANOVA followed by the Tukey’s post hoc comparison test. Different letters above the bars indicate significant differences (p < 0.05). (AU = Arbitrary Units) Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31029157), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Ca, Ch, ChHa, Eq, Ha, Md, Pm, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ca, Ch, ChHa, Eq, Ha, Md, Pm, Rb

Applications:

, , , , , , , , , , , , , , , ,

Applications: , , , , , , , , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB400-105AF350
Alexa Fluor 405	NB400-105AF405
Alexa Fluor 488	NB400-105AF488
Alexa Fluor 532	NB400-105AF532
Alexa Fluor 594	NB400-105AF594
Alexa Fluor 647	NB400-105AF647
Alexa Fluor 700	NB400-105AF700
Alexa Fluor 750	NB400-105AF750
Allophycocyanin	NB400-105APC
Allophycocyanin/Cy7	NB400-105APCCY7
Biotin	NB400-105B
DyLight 350	NB400-105UV
DyLight 405	NB400-105V
DyLight 488	NB400-105G
DyLight 550	NB400-105R
DyLight 594	NB400-105DL594
DyLight 650	NB400-105C
DyLight 680	NB400-105FR
DyLight 755	NB400-105IR
FITC	NB400-105F
HRP	NB400-105H
Janelia Fluor 549	NB400-105JF549
Janelia Fluor 646	NB400-105JF646
PE	NB400-105PE
PE/Atto594	NB400-105PEATT594
PE/Cy5.5	NB400-105PECY55
PE/Cy7	NB400-105PECY7
PerCP	NB400-105PCP
mFluor Violet 450 SE	NB400-105MFV450
mFluor Violet 500 SE	NB400-105MFV500
mFluor Violet 610 SE	NB400-105MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB400-105SS

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AF933

ACE-2 Antibody [Unconjugated]

(1 Review)

(279 Publications)

100 μg

AF933

Western blot shows lysates of human kidney tissue, mouse kidney tissue, and rat kidney tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse/Rat/Hamster ACE-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF933) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for ACE-2 at approximately 100 and 110 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

HEK293 human embryonic kidney cell line transfected with (A) human ACE-2 or (B) irrelevant protein, and eGFP was stained with Goat Anti-Human/Mouse/Rat/Hamster ACE-2 Affinity Purified Polyclonal Antibody (Catalog # AF933) followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (F0108). Quadrant markers were set based on Goat IgG control antibody (AB-108-C, data not shown). Staining was performed using our Staining Membrane-associated Proteins protocol.

ACE‑2 was detected in immersion fixed paraffin-embedded sections of human kidney using Goat Anti-Human ACE‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF933) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

ACE‑2 was detected in immersion fixed paraffin-embedded sections of hamster lung using Goat Anti-Human/Mouse/Rat/Hamster ACE‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF933) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to respiratory bronchioles. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

ACE‑2 was detected in immersion fixed frozen sections of rat lung using Goat Anti-Human/Mouse/Rat/Hamster ACE‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF933) at 10 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell surface in eputhelial cells in bronchioles. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

ACE‑2 was detected in immersion fixed paraffin-embedded sections of rat lung using Goat Anti-Human/Mouse/Rat/Hamster ACE‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF933) at 10 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

ACE‑2 was detected in immersion fixed paraffin-embedded sections of rat kidney using Goat Anti-Human/Mouse/Rat/Hamster ACE‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF933) at 10 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell surface in convoluted tubules. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Simple Western lane view shows lysates of human kidney tissue, loaded at 0.2 mg/mL. A specific band was detected for ACE‑2 at approximately 155 kDa (as indicated) using 10 µg/mL of Goat Anti-Human ACE‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF933) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

(A) Transduction of pLV pseudotyped with VSV-G (A,C) or CoV-2 Spike glycoprotein (B,D) in HEK293T (A,B) or Vero E6 (C,D) cells. The lentiviral backbone incorporates enhanced green fluorescent protein (eGFP) that is expressed upon integration into target cells. The fluorescence was recorded at 48 h post transduction. Magnification 4X. (E) Transduction efficiency of pLV pseudotyped with CoV-2 Spike glycoprotein in Vero E6, hACE2-HEK293T and 293T cells. The fluorescence was recorded at 48 h post transduction. The experiments were done in triplicates and standard error of mean was plotted as error bars. (F) Whole cell lysates from Vero E6, hACE2-293T and 293T cells were run on SDS-PAGE and probed with anti ACE2 antibody. Beta-actin was used as a loading control. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33154514), licensed under a CC-BY license. Not internally tested by R&D Systems.

Immunoblot analysis of ACE2 protein in the media from high glucose- or Ang II-stimulated mouse PT cells.(A) Mouse PT cells were incubated for 72 hrs in normal media (C, 7.8 mM D-glucose), with or without Ang II (10−7 M), high D-glucose (D-G, 25 mM), or high L-glucose (25 mM). Above graph is representative immunoblot for ACE2 in the media, showing bands at ∼90 kDa and ∼70 kDa. (B) Graphical representation of densitometry analysis of two ACE2 bands on immunoblots. For the ∼90 kDa band, *p<0.05 vs C, **p<0.001 vs C, **p<0.003 vs L-G; n = 5. For the ∼70 kDa band, *p<0.04 vs C; **p<0.001 vs C or L-G, **p<0.03 vs Ang II; n = 5. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0085958), licensed under a CC-BY license. Not internally tested by R&D Systems.

$Deglycosylation of ACE2 protein in media and cell lysates from mouse PT cells.Representative immunoblot for ACE2 treated without (−) or with (+) deglycosylation with PNGase F in the media (Lanes 1–2) and cell lysates (Lanes 3–4). Lanes 1 and 3: wildtype PT cells, Lanes 2 and 4: ACE2 knockout (KO) PT cells transfected with a human ACE2 vector, Lane 5: mouse kidney cortex. Lanes 1+ and 2+ show a reduction in the sizes of ACE2 fragments in media fractions to ∼75 kDa and ∼60 kDa for mouse ACE2, and to ∼80 kDa and ∼65 kDa for human ACE2, respectively. Lanes 3+ and 4+ show a reduction in the sizes of ACE2 in cell lysates to ∼85 kDa for both mouse and human ACE2 treated with the PNGase F, respectively. Lane 5+ shows a reduction in size of ACE2 in mouse cortex from ∼100 kDa to ∼85 kDa after treatment with PNGase F. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0085958), licensed under a CC-BY license. Not internally tested by R&D Systems.$

Immunoblot analysis of ACE2 protein in media and cell lysates from mouse PT cells.(A) Representative immunoblot for ACE2 protein in concentrated media (Lanes 1–3) and cell lysates (Lanes 4–6) from mouse PT cells. Lanes 1 and 4: wildtype cells, Lanes 2 and 5: ACE2 knockout (KO) cells, Lanes 3 and 6: ACE2 KO cells transfected with a human ACE2 expression vector, Lane 7: mouse kidney cortex showing a band at ∼100 kDa, used as a positive control. Lane 1 shows two bands in the media at ∼90 kDa and ∼70 kDa for mouse ACE2. Lane 3 shows two bands in the media for human ACE2 in transfected cells, at ∼110 kDa and ∼95 kDa. Lanes 4 and 6 show a single band in cell lysates at ∼100 kDa for mouse ACE2, and ∼120 kDa for human ACE2, respectively. Lanes 2 and 5 show no ACE2 bands detected on immunoblots of both media and cell lysates from untransfected ACE2 KO cells. (B) Increased ACE2 activity in the media from ACE2 KO cells transfected with a human ACE2 expression vector (HA-hACE2, 3.75 µg on 35 mm culture dishes). Untransfected cells and cells transfected with an empty pcDNA3 vector had no detectable ACE2 activity in the media. Numbers in parentheses represent mean values for ACE2 activity. *P<0.001 vs untransfected control or empty pcDNA3 vector, n = 4. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0085958), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu, Mu, Rt, Ha

Species Glossary

Reactivity: Hu, Mu, Rt, Ha

Applications:

, , , , ,

Applications: , , , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	FAB933U-100UG
Alexa Fluor 405	FAB933V-100UG
Alexa Fluor 488	FAB933G-100UG
Alexa Fluor 594	FAB933T-100UG
Alexa Fluor 647	FAB933R-100UG
Alexa Fluor 700	FAB933N-100UG
Alexa Fluor 750	FAB933S-100UG
Allophycocyanin	FAB933A-100
Biotin	BAF933
Phycoerythrin	FAB933P-100

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF933-SP
Carrier Free	FAB933V-100UG
Carrier Free	FAB933U-100UG
Carrier Free	FAB933T-100UG
Carrier Free	FAB933S-100UG
Carrier Free	FAB933R-100UG
Carrier Free	FAB933N-100UG
Carrier Free	FAB933G-100UG
Carrier Free	FAB933P-100
Carrier Free	FAB933A-100

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AF1062

PDGFR alpha Antibody [Unconjugated]

(4 Reviews)

(275 Publications)

100 μg

AF1062

Western blot shows lysates of mouse uterus tissue and mouse lung tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse PDGF Ra Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1062) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). Specific bands were detected for PDGF Ra at approximately 160-200 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

PDGF Ra was detected in immersion fixed frozen sections of mouse embryo using Goat Anti-Mouse PDGF Ra Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1062) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

PDGF Ra was detected in immersion fixed frozen sections of mouse embryo using 15 µg/mL Goat Anti-Mouse PDGF Ra Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1062) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the plasma membrane of mesenchymal cells. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Recombinant Human PDGF-AA (221-AA) stimulates proliferation in the NR6R-3T3 mouse fibroblast cell line in a dose-dependent manner (orange line), as measured by Resazurin (AR002. Proliferation elicited by Recombinant Human PDGF-AA (250 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse PDGF Ra Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1062). The ND50 is typically 0.2-1.6 µg/mL.

Extracellularly applied recombinant human alpha -syn PFFs induced cytoplasmic alpha -syn-immunoreactive inclusions in primary BCAS1(+) cell cultures. Immunostaining of oligodendroglial cells incubated with 1 μM alpha -syn PFFs for 24 h from days 3 (upper) and 4 (lower) after differentiation induction showing the ubiquitous development of thioflavin S-labeled inclusions in PDGFR alpha (+) cells and BCAS1(+) cells. In contrast, few BCAS1(−)/MBP(+) cells developed thioflavin S-labeled inclusions. Scale bar = 50 μm. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32727582), licensed under a CC-BY licence.

Rspo3 mRNAs are localized on telopodes that extend away from the cell bodies of the mouse VTTs. VTTs are marked by Lgr5 mRNA (red dots), Rspo3 mRNA (green dots) is localized away from the cell body, PDGFRa antibody mark VTTs cell bodies and telopodes. Scale bar–10 µm, in inset, green arrows point to Rspo3 mRNAs (green dots) localized on PDGFRa telopodes (blue). Telocyte cell body is marked by white dashed line. inset Scale bar–5 µm. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32321913), licensed under a CC-BY licence.

Leptin promotes OPC proliferation. Representative image of cultured OPCs stained with antibodies against LepRb (green) and PDGFR alpha (red). Scale bar: 25 μm. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep40397), licensed under a CC-BY licence.

LPC injection does not enhance leptin expression in the CNS. Representative images of LepRb (green) expression in combination with PDGFR alpha , GFAP NeuN, and CD11b (red) in the mouse spinal cord with or without LPC injection. Spinal cord sections were obtained 3 days after LPC injection. Graph indicates the relative intensity of leptin protein expression in indicated cell type (n = 3). P = 0.287452 (PDGFR alpha ), 0.181059 (GFAP), 0.199972 (NeuN), Student’s t-test, n.s. indicates no significant difference. *P 4-001-A, open histogram) followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). View our protocol for Staining Membrane-associated Proteins.

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

, , ,

Applications: , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 488	FAB1062G
Allophycocyanin	FAB1062A
Biotin	BAF1062
Phycoerythrin	FAB1062P

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	FAB1062P
Carrier Free	FAB1062A
Carrier Free	FAB1062G
Carrier Free	AF1062-SP

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NB400-104

SR-BI Antibody - BSA Free

(5 Reviews)

(227 Publications)

0.1 ml

NB400-104

Immunocytochemistry/Immunofluorescence: SR-BI Antibody [NB400-104] - SR-BI antibody was tested in human fibroblast samples fixed in 4% PFA and permeabilized in PBS (0.2% Tween). Primary incubation overnight at 4C using a 1:100 dilution in PBS (0.1% Tween) with 1% BSA. Secondary antibody is anti-rabbit conjugate to Alexa Fluor 488. SR-BI is shown in green and nucleus in blue (Hoescht 33342 stain).

Flow Cytometry: SR-BI Antibody [NB400-104] - An intracellular stain was performed on HeLa cells with SR-BI antibody NB400-104AF488 (blue) and a matched isotype control NBP2-24893AF488 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

Western Blot: SR-BI Antibody [NB400-104] - Detection of SR-BI in rat H4IIE total cell lysates and plasma membrane proteins. Photo courtesy of product review by verified customer.

Immunohistochemistry: SR-BI Antibody [NB400-104] - Immunolocalization of SR-BI in adult mink testis using NB400-104. SR-BI labeling is visible at the surface and along the outline of the large vacuole. Photo courtesy of R.M. Pelletier, University of Montreal.

Immunohistochemistry-Paraffin: SR-BI Antibody [NB400-104] - FFPE tissue section of mouse liver using SR-BI antibody (Lot 8310) at 1:300 dilution with HRP-DAB detection and hematoxylin counterstaining. The antibody generated a specific membrane signal of SR-BI protein in the murine hepatocytes.

<b>Genetic Strategies Validation. </b>Knockdown Validated: SR-BI Antibody [NB400-104] - HCV replication was increased by CD36. Silence of CD36 decreased HCV infection and replication (n = 3). Huh7.5 cells were transfected with 0.2 u g CD36-HA plasmid or with 150 pmol siRNA for CD36 (B) for 48 hrs, and then were washed and infected with HCV (150 IU/cell) for 2 hrs, followed by washing and continuously incubating. Intracellular HCV RNA and proteins were detected in 96 hrs, and the cytotoxicity of siRNAs was measured in 96-wells plate with a MTT assay in B. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep21808) licensed under a CC-BY license.

Immunohistochemistry-Paraffin: SR-BI Antibody [NB400-104] - Staining of human adrenal cortex.

Immunocytochemistry/Immunofluorescence: SR-BI Antibody [NB400-104] - HeLa cells were fixed and permeabilized for 10 minutes using -20C MeOH. The cells were incubated with anti-SR-BI at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry-Paraffin: SR-BI Antibody [NB400-104] - FFPE tissue section of mouse liver using SR-BI antibody (Lot R-4) at 1:300 dilution with HRP-DAB detection and hematoxylin counterstaining. The antibody generated mainly a membranous signal of SR-BI protein in the murine hepatocytes.

Flow Cytometry: SR-BI Antibody [NB400-104] - Analysis of Huh7 and HepG2 cells using SR-B1 antibody NB400-104. Courtesy of Bruno Sainz, Jr., PhD, University of Illinois at Chicago.

Immunofluorescence assay on GIM biopsies of HCV-transplanted patients.Immunofluorescence assay on GIM polyp biopsies of HCV-transplanted patients, using antibodies against HCV receptors: CD81, SR-B1, Claudin-1, Occludin (Original magnification X400). (A, E) CD81 in colon (patient 26); (B, F) Claudin-1 in colon (patient 26); (C, G) Occludin in colon (C patient 28, G patient 26); (D) SR-B1 in antrum (patient 28); (H) SR-B1 in colon (patient 26). All the sections were clearly positive for the analyzed HCV receptors before (A-D) and after transplantation (E-H). Negative controls (I-N) were performed on GIM biopsies of HCV positive patients by omitting the primary antibodies, and by using polyclonal FITC-conjugated Donkey anti-mouse (I) and anti-rabbit (L, M, N) as secondary antibodies. (I CD81 in antrum; L Claudin-1 in polyp colon; M Occludin in antrum; N SR-B1 in polyp colon. Nuclei were counterstained with DAPI (blue). Scale bar 50 um. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0181683), licensed under a CC-BY licence.

HCV replication was increased by CD36. (B) Silence of CD36 decreased HCV infection and replication (n = 3). Huh7.5 cells were transfected with 0.2 ug CD36-HA plasmid (A) or with 150 pmol siRNA for CD36 (B) for 48 hrs, and then were washed and infected with HCV (150 IU/cell) for 2 hrs, followed by washing and continuously incubating. Intracellular HCV RNA and proteins were detected in 96 hrs, and the cytotoxicity of siRNAs was measured in 96-wells plate with a MTT assay in B. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep21808), licensed under a CC-BY licence.

ALDH2 rs671 modulates HDL-C levels in mice and human liver through increasing poly(ADP-ribosyl)ation of LXRalpha due to attenuated ALDH2/PARP1 interaction. (C) Western blotting analysis of ABCA1, ALDH2, LXRalpha, and SR-B1 expression in mouse liver tissue. WT, n = 3; rs671, n = 3. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35393951), licensed under a CC-BY licence.

ZA treatment increases ABCA1 and apoA-1 expression.(a) Western blot analysis of ABCA1, ABCG1 and SR-BI expression in untreated (ZA-) and ZA-treated (ZA+) THP-1 cells, U937 cells, CD14+ cells and corresponding DC subsets (DCTHP1, DCU937, DC). ABCA1 was already detectable in DCU937 and DC and its expression was upregulated by ZA. beta-tubulin was employed to check the equal protein loading per lane. The results are obtained from one representative experiment of three experiments. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms15663), licensed under a CC-BY licence.

Differentiation and characteristics of iHLCs. (d) Western blot analysis of protein expression in HCV-infected iHLCs compared to Huh7.5 cells of different host factors crucial for HCV infection. Tubulin served as loading control. Full-length blots are presented in Supplementary Figure 1. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29497123), licensed under a CC-BY licence.

Expression of cellular HCV receptors at the mRNA and protein levels.(a) mRNA copy numbers of CD81, occluding (OCLN), claudin-1 (CLDN1) and SR-BI were quantified in OASF, RASF and Huh-7.5 cells, normalized to the reference gene GAPDH. (b) Detection of CD81 on Huh-7.5 cells, OASF, and RASF by FACS analysis. (c-d) Detection of OCLN, CLDN1 and SR-BI in OASF and RASF by immunoblot analysis. Huh-7.5 cells were used as positive control and Hep-56.1D as negative control. In both the real-time PCR and the FACS analysis experiments the number of biological replicates was 3 and the number of experimental replicates was 2 (total n = 6). In immunoblotting detection, the number of biological replicates was 2 and the number of experimental replicates was 2 (n = 4). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26643193), licensed under a CC-BY licence.

miR-27a/b inhibit PINK1 accumulation upon mitochondrial damage. a, b Overexpression of miR-27a/b inhibited PINK1 accumulation upon mitochondrial damage. 48 h post-transfection, HeLa cells were treated with 10 μM CCCP or combination of 10 μM oligomycin and 4 μM antimycin as indicated for 2 h (n = 4, two-way ANOVA). c, d Inhibition of endogenous miR-27a/b increased PINK1 accumulation upon CCCP treatment. (n = 4, two-way ANOVA). PINK1 levels were normalized to corresponding GAPDH level and quantified as a percentage of control. Values are mean ± SEM (n.s. = non-significant, *p < 0.05, **p < 0.01, ***p < 0.001) Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27456084), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

A beta induces the expression of cell cycle regulatory proteins. (A) SK-N-MC cells were exposed to A beta (5 μM) for 24 h. The mRNA expression levels of CDK5, P35 and P39 were analyzed by real-time PCR. The mRNA expression level was normalized by beta -actin mRNA expression level. Data represent the mean ± SE. n = 4. (B)hif1 alpha specific- and non-targeting (NT) siRNA were transfected to the cells for 24 h prior to A beta treatment. Cyclin D1, CDK4, cyclin E, CDK2, HIF1 alpha and beta -actin was detected by western blot. n = 3. (C–F) Cells were pretreated with trehalose (10 μM), rapamycin (10 nM), PF4708671 (10 μM) and cycloheximide (4 μM) for 30 min prior to A beta treatment for 24 h. Cyclin D1, CDK4, cyclin E, CDK2 and beta -actin were detected by western blot. n = 3–6. (G) Mouse hippocampal neurons were transfected with hif1 alpha specific- and NT siRNAs for 24 h prior to A beta treatment for 24 h. Samples were blotted with Cyclin D1, CDK4, cyclin E, CDK2 and beta -actin specific antibodies. n = 3–6. (H) Mouse hippocampal neurons were pretreated with trehalose (10 μM) for 30 min and incubated with A beta for 24 h. cyclin D1, CDK4, cyclin E, CDK2, HIF1 alpha and beta -actin were analyzed by western blot. n = 3–6. Data are presented as a mean ± SE. *p < 0.05 vs. control, #p < 0.05 vs. A beta treatment. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28790888), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Yap deficiency suppresses cell proliferation in vivo and in vitro. (A-F) The Ki67 positive ratio of lens epithelial cells decreased in Yap-deficient mice at different stages (arrowheads indicate Ki67 positive cells). (G) The relative number of Ki67 positive lens epithelial cells (number of Ki67 positive lens epithelial cells / lens epithelium area). The data are shown as mean ± S.E.M. (Student’s t-test, *P<0.05, **P<0.01, n=10). (H-I) Knockdown efficiency of Yap in alpha TN4 cell using siRNA. (J-K) Cell viability and growth assay revealed that proliferation was downregulated in Yap knockdown alpha TN4 cells. The data are shown as mean ± S.E.M. (Two-way RM ANOVA, **P<0.01, n=5). Scale bars: 50 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31011480), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Knockdown of CIP2A inhibited cell proliferation and DNA synthesis of HPV-16E7-expressing cells(A) Western blot analysis of protein level of 16E7 and CIP2A in RPE1-16E7 cells and (B) with CIP2A siRNA for 48 hr. (C) CCK8 assay of cell proliferation of RPE1-16E7 cells with CIP2A siRNA. (D) Flow cytometry of cells with CIP2A siRNA and labeled with BrdU for 2 hr, then stained with PI and BrdU; and (E), Quantification. Babe, vector control. **, P < 0.01. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/25650660), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

ATO interferes with efficient HAdV replication center formation and reorganization of PML‐NBs. H1299 cells were infected with HAdV‐C5 wt at a multiplicity of 20 FFU per cell, treated with the depicted concentrations of ATO at 2 h p.i., fixed 48 h p.i. with 4% PFA and double labeled with mAb B6‐8 ( alpha ‐E2A) and pAb NB100‐59787 ( alpha ‐PML). Primary antibodies were detected using Alexa488 (PML, green) and Alexa647 (E2A, red) conjugated secondary antibodies. A) Number of PML‐NBs per cell in uninfected cells was determined using Volocity for at least n = 461 cells from two independent biological replicates. Statistically significant differences were determined using a one‐way ANOVA and Dunnet's T3 test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001. B) The proportion of infected cells showing track‐like redistribution of PML‐NBs was determined by counting for at least n = 212 cells and normalization to untreated infected cells. C) The proportion of infected cells showing formation of HAdV replication centers marked by the viral protein E2A (lower plot) was determined by counting for at least n = 212 cells and normalization to untreated infected cells. D) Cells showing either viral replication centers with PML track‐like structures, replication centers without PML track‐like structures, no replication centers but PML track‐like structures or no replication centers, and no PML track‐like structures were counted for at least n = 214 (virus infected cells due to E2A signal detected: either untreated/0 µm or treated with ATO/1 or 2 µm) and represented in pie charts. Statistically significant differences were determined using a one‐way ANOVA and Dunnet's T3 test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001. E) H1299 cells were infected with HAdV‐C5 wt at a multiplicity of 20 FFU per cell, and treated with 0 or 2 µm of ATO at 2 h p.i. After 48 h, the cells were fixed with 4% PFA and stained using pAb NB100‐59787 ( alpha ‐PML) and mAb 6A‐11 ( alpha ‐E4orf3). Primary antibodies were detected using Alexa488 (PML, green) and Alexa647 (E4orf3, red) coupled secondary antibodies. Representative staining patterns for at least 30 uninfected cells treated with 0 or 2 µm ATO are shown in panels (a)–(d) and (m)–(p), infected cells treated with 0 or 2 µm ATO are shown in panels (e)–(l) and (q)–(x). Overlays of single fluorescence pictures (merge) are shown in panels (d), (h), (l), (p), (t), and (x). Data corresponds to two independent biological replicates performed and counted by different operators to avoid operator bias. Scale bar represents 10 µm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32328411), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$ABCC4 protein expression level in CRC cell lines. Western blot performed in standard reducing SDS PAGE conditions using goat anti ABCC4 (#PA5 18315, Thermo Scientific) and rabbit anti ABCG2 (#ORB 155559 Biorbyt). (A) Protein expression level of ABCC4 and ABCG2 in HT-29 stably overexpressing transcription factor Snail (HT29/Snail) and control HT-29. ABCC4 level in the membrane fraction (obtained by biotynylation using EZ-Link Sulfo -NHS-Biotin Thermo Scientific kit) of HT-29 control cells and HT-29 Snail n = 3. (B) ABCC4 protein expression level in CRC cells in different states of EMT: CCD841CoN (most epithelial), CaCo-2 (moderate EMT), and Colo-320 (most mesenchymal) n = 3. (C) ABCC4 protein abundance in Extracellular Vesicles (EVs) released from HT-29 control cells and two HT-29 stably overexpressing transcription factor Snail clones (HT-29/Snail and HT-29/Snail17), n = 2. (D) Intracellular cAMP level measurement. Accumulation of cAMP in HT29 cells was measured using a cAMP competitive kit (#581001 Cayman Chemical). Cells were incubated for 24 h with MK571 20 µM, or untreated ones were assayed according to the manufacturer’s protocol. Calculation were conducted using the Cayman data sheet. cAMP concentration of HT29 was set as 100%. T-test performed, n = 5; * p < 0.05; ** p < 0.005; *** p < 0.001. NS—not statistically significant. (E) PKA phosphorylation profile analysis. HT29 Snail cells were seeded on a 6-well plate. Then, 24 h after, full growth medium was changed into starving (FBS free) medium for 24 h. Next, 20uM of MK571 was added to cells for 60, 30, 5, and 1 min. Cells without the starving procedure were used as a positive control, and negative control cells were not treated with MK571. Phosphorylation profile analysis was performed using phospho-(ser/thr) PKA Substrate Antibody #9621 (Cell Signaling Technology). Significant time- (exposure) related impact on the phosphorylation profile was observed for 42 kDa and 95–100 kDa proteins in HT29 Snail cells compared to no time-related changes in control cells, n = 3. (F) HT-29/Snail PKA phosphorylation profile analyzed with densitometry; statistical significance estimated using T-test. * p < 0.05; ** p < 0.005; *** p < 0.001. NS—not statistically significant. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/33261018), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Reactivity:

Hu, Mu, Rt, Po, ChHa, Ha, Md, Pm, Rb, SyHa

Species Glossary

Reactivity: Hu, Mu, Rt, Po, ChHa, Ha, Md, Pm, Rb, SyHa

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB400-104AF350
Alexa Fluor 405	NB400-104AF405
Alexa Fluor 488	NB400-104AF488
Alexa Fluor 532	NB400-104AF532
Alexa Fluor 594	NB400-104AF594
Alexa Fluor 647	NB400-104AF647
Alexa Fluor 700	NB400-104AF700
Alexa Fluor 750	NB400-104AF750
Allophycocyanin	NB400-104APC
Biotin	NB400-104B
DyLight 350	NB400-104UV
DyLight 405	NB400-104V
DyLight 488	NB400-104G
DyLight 550	NB400-104R
DyLight 594	NB400-104DL594
DyLight 650	NB400-104C
DyLight 680	NB400-104FR
DyLight 755	NB400-104IR
FITC	NB400-104F
HRP	NB400-104H
Janelia Fluor 549	NB400-104JF549
Janelia Fluor 646	NB400-104JF646
PE	NB400-104PE
PerCP	NB400-104PCP
mFluor Violet 450 SE	NB400-104MFV450
mFluor Violet 500 SE	NB400-104MFV500
mFluor Violet 610 SE	NB400-104MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB400-104SS

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MAB1835

TGF beta 1/2/3 Antibody (1D11) [Unconjugated]

(1 Review)

(222 Publications)

100 μg

MAB1835

Recombinant Human TGF‑ beta 1 (Catalog # 240‑B) inhibits Recombinant Mouse IL‑4 (Catalog # 404-ML) induced proliferation in the HT‑2 mouse T cell line in a dose-dependent manner (orange line). Inhibition of Recombinant Mouse IL‑4 (7.5 ng/mL) activity elicited by Recombinant Human TGF‑ beta 1 (1 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-TGF‑ beta 1, 2, 3 Monoclonal Antibody (Catalog # MAB1835). The ND50 is typically0.25‑1.25 µg/mL.

Apoptotic cell-induced immunomodulation in mice with collagen-induced arthritis (CIA) is dependent on transforming growth factor (TGF)-beta . CIA mice that received or did not receive apoptotic cells, with or without anti-TGF-beta blocking antibody were scored daily (a). Data are shown as mean ± SEM of five mice per group from one of two representative experiments; *p < 0.05, **p < 0.01 (Friedman test analysis of variance (ANOVA) with Dunn's multiple comparisons test). Anti-collagen IgG2a antibodies (Ab) were quantified in plasma (b). Data from 5 to 16 mice from two to three independent experiments (no difference, Kruskal-Wallis test ANOVA with Dunn’s multiple comparisons test). Lymph node cells were collected, cultured, and T cell proliferation in response to collagen protein or CD3-specific Ab stimulations was evaluated (c). Data are shown as mean ± SEM from one of two representative experiments; *p < 0.05, **p < 0.01 (nonparametric unpaired t test). Percent and absolute number of Foxp3+ T regulatory cells (Treg) were evaluated in the spleen (d) and suppressive assays were performed by isolating and adding Treg at a different ratio into collagen-specific cultures and responder T cell proliferation was assessed (e). Data are shown as mean ± SEM from one experiment (d Kruskal-Wallis test ANOVA with Dunn’s multiple comparisons test; e Friedman test ANOVA with Dunn’s multiple comparisons test). Plasmacytoid dendritic cells (pDC), conventional dendritic cells (cDC) and macrophages (macro) were isolated and cultured with naïve allogenic T cells to determine Treg polarization (f), and their response to TLR ligand was assessed through evaluation of CD40 costimulatory molecule expression (g). Data from individual mouse plus mean (black bar) from one experiment representative of four are shown; *p < 0.05, ***p < 0.001 (f ordinary one-way ANOVA with Tukey’s multiple comparisons test; g Kruskal-Wallis test ANOVA with Dunn’s multiple comparisons test). BrDU 5-Bromo-2’-deoxyuridine Image collected and cropped by CiteAb from the following publication (//arthritis-research.biomedcentral.com/articles/10.1186/s13075-016-1084-0), licensed under a CC-BY license. Not internally tested by R&D Systems.

TGF beta 1 stimulates the expression of ECM components in human and rabbit primary cells. (A) Selected ECM mRNA transcripts were measured using qRT-PCR in human RPTEC or human IPF134 following 48 hr stimulation with TGF beta 1 (10 ng/mL). Data show transcript abundance relative to the unstimulated control (indicated by the dashed line) and are shown as mean ± SD of four (RPTEC) or seven (IPF134) independent experiments. ****P < 0.0001; ****P < 0.001; **P < 0.01; Student’s t-test. (B) The accumulation of ECM in human RPTEC (i) or human IPF (ii) cells stimulated with 10ng/mL TGF beta 1 was evaluated via measurement of the incorporation of 14C-labelled amino acids into the deposited ECM. Data presented are the mean ± SD of three independent experiments. **P < 0.01; Student’s t-test. (C) Increased mature ECM accumulation following treatment of cells with a pro-fibrotic stimulus can be visualised following in-situ fluorescent staining using the Flamingo dye. Cells from three different systems (primary human RPTEC; primary human IPF134; co-culture of primary rabbit RPTEC with primary rabbit renal fibroblasts) were cultured in the absence (top) or presence (bottom) of 10 ng/ml TGF beta 1 before decellularised matrix was fixed and stained in situ with Flamingo fluorescent dye. Images show a single field. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28855577), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

All-Multi

Species Glossary

Reactivity: All-Multi

Applications:

, ,

Applications: , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	FAB1835U
Alexa Fluor 405	FAB1835V
Alexa Fluor 488	FAB1835G
Alexa Fluor 532	FAB1835X
Alexa Fluor 594	FAB1835T
Alexa Fluor 647	FAB1835R
Alexa Fluor 700	FAB1835N
Alexa Fluor 750	FAB1835S
Allophycocyanin	FAB1835A
Biotin	FAB1835B
DyLight 350	FAB1835D
DyLight 405	FAB1835E
DyLight 488	FAB1835K
DyLight 550	FAB1835L
DyLight 594	FAB1835M
DyLight 650	FAB1835W
DyLight 680	FAB1835Y
DyLight 755	FAB1835Z
FITC	FAB1835F
HRP	FAB1835H
Janelia Fluor 549	FAB1835I
Janelia Fluor 646	FAB1835J
PerCP	FAB1835C
mFluor Violet 450 SE	FAB1835MFV450
mFluor Violet 500 SE	FAB1835MFV500
mFluor Violet 610 SE	FAB1835MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	MAB1835-SP
Carrier Free	MAB1835-500

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AF808

Osteopontin/OPN Antibody [Unconjugated]

(2 Reviews)

(201 Publications)

100 μg

AF808

Osteopontin/OPN was detected in immersion fixed C2C12 mouse myoblast cell line (left panel, positive stain) and mouse splenocytes (right panel, negative stain) using Goat Anti-Mouse Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF808) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and secreted molecule. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Osteopontin/OPN was detected in perfusion fixed frozen sections of mouse thymus using Mouse Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF808) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Recombinant Mouse Osteopontin/OPN (Catalog # 441-OP), immobilized onto a microplate, supports the adhesion of the HEK293 human embryonic kidney cell line in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Mouse Osteopontin/OPN (2 µg/mL) is neutralized (green line) by increasing concentrations of Mouse Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF808). The ND50 is typically 1‑3 µg/mL.

Notch ligands Jag1 and Dll1 are both required for segregation of hepatocytic fate centrally and biliary fate peripherally.(A) Immunolabeling for OPN and HNF4A of BMEL cells presented with DLL4 on 30 kPa substrates. Control cells were transduced with an shRNA vector coding for a non-mammalian target. shJag1 and shDll1 cells were transduced with shRNA vectors targeting Jag1 and Dll1, respectively. Scale bar is 150 µm. (B) Confocal imaging of immunolabeled SOX9 and HNF4A in control, shJag1, and shDll1 cells presented with DLL4 on 30 kPa substrates. Scale bar is 75 µm. (C) Quantification of OPN+ cell counts of control, shJag1, and shDll1 cells presented with DLL4 on 30 kPa substrates. (D) Quantification of SOX9 and HNF4A intensity of control, shJag1, and shDll1 cells presented with DLL4 on 30 kPa substrates. (C, D) Mean ± 95% CI.10.7554/eLife.38536.024Figure 7—source data 1.Summary table for OPN data in Figure 7C.10.7554/eLife.38536.025Figure 7—source data 2.Summary table for SOX9 and HNF4A data in Figure 7D.Summary table for OPN data in Figure 7C.Summary table for SOX9 and HNF4A data in Figure 7D.Regression analysis of OPN+ cell counts.Data in Figure 7B were separated into peripheral and central subsets for which dimensionless radius was greater than 0.75 (R>0.75) and less than 0.75 (R0.75) and less than 0.75 (R0.75) and less than 0.75 (R0.9) compared with central intensity (R<0.1) using Welch’s t-test.Immunolabeling of OPN and CK19 at t=24h and cell density with radius at t=72h.(A) Immunolabeling at t=24h of BMEL cells on 30 kPa substrates presented with IgG and DLL4 for the biliary markers OPN and CK19. Scale bar indicates 75 µm. (B) Measurement of cell density with radius for 30 kPa and 4 kPa substrates at t=72h. Vertical black bars indicate computed mean radius using the most central 95% of cells within the data set.Immunolabeling for OPN with 300, 600, and 1000 µm diameter patterns.(A) Immunolabeling for OPN of BMEL cells presented with DLL4 on 30 kPa substrates at t=72h. Both 300 µm and 1000 µm pattern diameters were included in this experiment in addition to the 600 µm pattern diameter. Scale bars indicate 150 µm. (B) Quantification of peak OPN+ cell counts for 300, 600, and 1000 µm diameter pattern. Boxplots show median, 25th and 75th percentiles (hinges), and 1.5 × IQR (whiskers). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30589410), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

, , , ,

Applications: , , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Biotin	BAF808
Phycoerythrin	IC808P

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	IC808P
Carrier Free	AF808-SP

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MAB208

CXCL8/IL-8 Antibody (6217) [Unconjugated]

(1 Review)

(180 Publications)

100 μg

MAB208

Western blot shows lysates of THP‑1 human acute monocytic leukemia cell line untreated (-) or treated (+) with 200 nM PMA and 10 ug/mL LPS for 24 hours and 3 hours. PVDF membrane was probed with 3 µg/mL of Mouse Anti-Human IL-8/CXCL8 Monoclonal Antibody (Catalog # MAB208) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for IL-8/CXCL8 at approximately 10 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

IL-8/CXCL8 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Mouse Anti-Human IL-8/CXCL8 Monoclonal Antibody (Catalog # MAB208) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Simple Western lane view shows human PBMC conditioned media untreated (-) or treated (+) with 1 μg/mL LPS for 24 hrs, loaded at 0.2 mg/mL. A specific band was detected for IL-8/CXCL8 at approximately 8 kDa (as indicated) using 50 µg/mL of Mouse Anti-Human IL-8/CXCL8 Monoclonal Antibody (Catalog # MAB208) . This experiment was conducted under reducing conditions and using the 2-40 kDa separation system.

Recombinant Human IL-8/CXCL8 (Catalog # 208-IL) chemoattracts the BaF3 mouse pro‑B cell line transfected with human CXCR2 in a dose-dependent manner (orange line). The amount of cells that migrated through to the lower chemotaxis chamber was measured by Resazurin (Catalog # AR002). Chemotaxis elicited by Recombinant Human IL-8/CXCL8 (20 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human IL-8/CXCL8 Monoclonal Antibody (Catalog # MAB208). The ND50 is typically0.08-0.4 µg/mL.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	IC208U-100UG
Alexa Fluor 350	FAB208U
Alexa Fluor 405	IC208V-100UG
Alexa Fluor 405	FAB208V
Alexa Fluor 488	IC208G-100UG
Alexa Fluor 488	FAB208G
Alexa Fluor 532	FAB208X
Alexa Fluor 594	IC208T-100UG
Alexa Fluor 594	FAB208T
Alexa Fluor 647	IC208R-100UG
Alexa Fluor 647	FAB208R
Alexa Fluor 700	IC208N-100UG
Alexa Fluor 700	FAB208N
Alexa Fluor 750	IC208S-100UG
Alexa Fluor 750	FAB208S
Allophycocyanin	FAB208A
Biotin	FAB208B
DyLight 350	FAB208D
DyLight 405	FAB208E
DyLight 488	FAB208K
DyLight 550	FAB208L
DyLight 594	FAB208M
DyLight 650	FAB208W
DyLight 680	FAB208Y
DyLight 755	FAB208Z
Fluorescein	IC208F
HRP	FAB208H
Janelia Fluor 549	FAB208I
Janelia Fluor 646	FAB208J
PerCP	FAB208C
Phycoerythrin	IC208P
mFluor Violet 450 SE	FAB208MFV450
mFluor Violet 500 SE	FAB208MFV500
mFluor Violet 610 SE	FAB208MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	IC208F
Carrier Free	IC208P
Carrier Free	MAB208-SP
Carrier Free	MAB208-500
Carrier Free	IC208V-100UG
Carrier Free	IC208U-100UG
Carrier Free	IC208T-100UG
Carrier Free	IC208S-100UG
Carrier Free	IC208R-100UG
Carrier Free	IC208N-100UG
Carrier Free	IC208G-100UG

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NBP2-24729

TLR9 Antibody (26C593.2) - BSA Free

(4 Reviews)

(152 Publications)

0.1 mg

NBP2-24729

<b>Genetic Strategies Validation. </b>Knockdown Validated: TLR9 Antibody (26C593.2) [NBP2-24729] - Expression of TLR-9 protein in HB cells before and after 48 hr of transfection. Beta-actin was used as an internal control. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0092748) licensed under a CC-BY license.

Flow Cytometry: TLR9 Antibody (26C593.2) [NBP2-24729] - Analysis of colonic LP or peritoneal cavity (PerC) of normal mice were evaluated by flow cytometry. B cells were intracellularly stained with the anti-TLR9 antibody after the cell surface staining with anti-B220 and CD5 antibodies, and examined using flow cytometry. N = 3, performed twice. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0146191) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: TLR9 Antibody (26C593.2) [NBP2-24729] - Cbl-b is required for the endocytic transit of TLR9. Representative confocal microscopic images of splenocytes from mice with indicated genotypes. For experiments, cells were stimulated through the BCR (green) for 30 minutes then fixed and stained for TLR9 (blue) and Lamp-1 (red)(n = 3). Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0089792) licensed under a CC-BY license.

Immunohistochemistry-Paraffin: TLR9 Antibody (26C593.2) [NBP2-24729] - Monkey retina tissue. Image from verified customer review.

Flow Cytometry: TLR9 Antibody (26C593.2) [NBP2-24729] - Gating strategy for the detection of TLR-7 and -9 in B cell subsets. PBMC were isolated from whole blood and stained for surface markers before cells were fixed, permeabilised and stained for TLR-9. FSC and SSC were first used to gate out debris and SSC-A and SSC-H was utilized to eliminate duplicates. Further gating was done on CD45 and CD19, to target B cells. To separate between the different B cell populations, we gated on CD27 and IgD (D), followed by TLR-9 expression on these subsets. Data from one representative patient is shown. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0120383) licensed under a CC-BY license.

Western Blot: TLR9 Antibody (26C593.2) [NBP2-24729] - Analysis of TLR9 in A) human PBMC, B) human intestine, C) mouse intestine, and D) rat intestine tissue lysates using this antibody at a dilution of 3 ug/ml.

Flow Cytometry: TLR9 Antibody (26C593.2) [NBP2-24729] - Expression of TLR9 on turbinate epithelial cells from healthy controls compared to turbinate and polyp epithelial cells from patients with CRSwNP, n = 5 (A). Intracellular staining for TLR9 (open histogram, black line) and isotype control (filled histogram) on turbinate epithelial cells from a healthy control (B), turbinate epithelial cells (C) and polyp epithelial cells from a patient (D), analysed using flow cytometry. Results are presented as mean +/- SEM, **P<0.01. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0105618) licensed under a CC-BY license.

TLR9 activation upregulated PD-L1 expression by promoting STAT3 Tyr705 phosphorylation in HCC cells. (A) PD-L1 protein expression after treatment with ODN2216 with different concentrations or the indicated times (different concentrations: 0, 5, 10, or 20uM; times: 0, 6, 12, 24, and 36 hours in 10 uM) in Hep3B cells. PD-L1 protein levels were analyzed by Western blotting. (B) PD-L1 protein expression after treatment with ODN2216 with different concentrations or the indicated times (different concentrations: 0, 2.5, 5, or 10uM; times: 0, 6, 12, 24, and 36 hours in 10 uM) in Huh7 cells. PD-L1 protein levels were analyzed by Western blotting. (C) PD-L1 protein expression after treatment with ODN1585 with different concentrations or the indicated times (different concentrations: 0, 2.5, 5, and 10 uM; times: 0, 6, 12, 24, and 36 hours in 5uM) in Hepa1-6 cells. PD-L1 protein levels were analyzed by Western blotting. (D) PD-L1+ tumor cells were detected by flow cytometry after TLR9 agonist (Hep3B and Huh7 cells with ODN2216; Hepa1-6 cells with ODN1585) treatment with indicated concentration. (values are mean +/- SD, *p < 0.05, **p < 0.01, ***p<0.001, NS indicates no significance). (E) PD-L1 expression after TLR9 overexpression in Huh7 cells. PD-L1 expression levels were analyzed by immunofluorescence. (F) PD-L1 protein expression after TLR9 overexpression in Huh7 cells. PD-L1 protein levels were analyzed by Western blotting. (G) PD-L1 protein expression in TLR9 knockdown or TLR9 rescue Hep3B cells. PD-L1 protein levels were analyzed by Western blotting. (H and I) mRNA levels of PD-L1 in Hep3B (H) and Huh7 (I) cells measured by qRT-PCR after stimulation with different concentrations of ODN2216. (values are mean +/- SD, *p < 0.05, **p < 0.01, NS indicates no significance) (J) TLR9 overexpression-induced PD-L1 expression after MYC, JUN, IRF1, IRF3, STAT1 or STAT3 silencing. PD-L1 mRNA expression in Huh7 cells was analyzed after TLR9 overexpression alone or in the presence of MYC-, JUN-, IRF1-, IRF3-, STAT1- or STAT3-specific siRNA or siRNA-NC. (values are mean +/- SD, ***p<0.001). (K) p-STAT3 (Tyr705) levels in Hep3B cells after treatment with different concentrations of ODN2216 (a TLR9 agonist; ODN2216: 0, 2.5, 5, or 10uM) analyzed by Western blotting. (L) p-STAT3 (Tyr705) levels after TLR9 overexpression in Huh7 cells analyzed by Western blotting. (M) TLR9 overexpression-induced p-STAT3 (Tyr705) levels after TLR9 inhibition. p-STAT3 (Tyr705) levels were analyzed after TLR9 overexpression alone or in the presence of the TLR9 antagonist chloroquine diphosphate. (N) p-STAT3-induced PD-L1 levels after STAT3 inhibition. PD-L1 levels were analyzed after TLR9 overexpression alone or in the presence of the STAT3-specific small molecular inhibitor BP-1-102. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32483468), licensed under a CC-BY licence.

Reactivity:

Hu, Mu, Rt, Ca, Eq, Pm, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, Ca, Eq, Pm, Pm

Applications:

, , , , , , , , , , ,

Applications: , , , , , , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Alt Catalog#

IMG-305A

Alt Catalog# IMG-305A

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP2-24729AF350
Alexa Fluor 405	NBP2-24729AF405
Alexa Fluor 488	NBP2-24729AF488
Alexa Fluor 532	NBP2-24729AF532
Alexa Fluor 594	NBP2-24729AF594
Alexa Fluor 647	NBP2-24729AF647
Alexa Fluor 700	NBP2-24729AF700
Alexa Fluor 750	NBP2-24729AF750
Allophycocyanin	NBP2-24729APC
Biotin	NBP2-24729B
Biotin	NBP2-24890
DyLight 350	NBP2-24729UV
DyLight 405	NBP2-24729V
DyLight 488	NBP2-24729G
DyLight 550	NBP2-24729R
DyLight 594	NBP2-24729DL594
DyLight 650	NBP2-24729C
DyLight 680	NBP2-24729FR
DyLight 755	NBP2-24729IR
FITC	NBP2-24729F
HRP	NBP2-24729H
Janelia Fluor 549	NBP2-24729JF549
Janelia Fluor 646	NBP2-24729JF646
PE	NBP2-24907
PE	NBP2-24729PE
PerCP	NBP2-24729PCP
mFluor Violet 450 SE	NBP2-24729MFV450
mFluor Violet 500 SE	NBP2-24729MFV500
mFluor Violet 610 SE	NBP2-24729MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Azide Free	NBP2-24863
BSA Free	NBP2-24729SS

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NB100-56566

TLR4 Antibody (76B357.1) - BSA Free

(5 Reviews)

(138 Publications)

0.1 mg

NB100-56566

Immunohistochemistry: TLR4 Antibody (76B357.1) [NB100-56566] - Pericryptal Myofibroblasts are Responsible for Increased TLR4 Expression in a Subset of CRCs. Double-stained immunofluorescence for TLR4 (green) and vimentin (red) in normal (I), adenoma (II), and colon adenocarcinoma (III) (10x). In the stromal compartment of CRCs, immunofluorescent staining for TLR4 localized to the pericryptal myofibroblasts in a subset of samples. Image collected and cropped by CiteAb from the following publication (//www.jeccr.com/content/33/1/45), licensed under a CC-BY license.

Western Blot: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis using 2 ug/mL on (A) human intestine and 6 ug/mL on (B) mouse intestine and (C) rat intestine lysate.

Flow (Intracellular): TLR4 Antibody (76B357.1) [NB100-56566] - Analysis using PE conjugate of NBP2-27149. An intracellular stain was performed on Jurkat cells with TLR4 antibody (76B357.1) NBP2-27149PE (blue) and an isotype control MAB004 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to phycoerythrin.

Immunocytochemistry/Immunofluorescence: TLR4 Antibody (76B357.1) [NB100-56566] - RH-30 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-TLR4 Antibody (76B357.1) NB100-56566 at 1 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Immunohistochemistry: TLR4 Antibody (76B357.1) [NB100-56566] - Immunofluorescent staining of TMAs. Representative tissue cores from normal (I), adenomatous polyps (II), and CRC (III and IV) are shown. Image collected and cropped by CiteAb from the following publication (//www.jeccr.com/content/33/1/45), licensed under a CC-BY license.

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis of rat salivary gland tissue section at 1:100 dilution. The antibody generated a membrane-cytoplasmic staining in the tissue with stronger signal in ductal epithelial cells.

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Tissue section of normal human skin stained with antibody at 5 ug/mL. Membrane-cytoplasmic immunopositivity of TLR4 was primarily observed in the pigmented basel cells and the adjacent keratinocytes in the epidermal layer.

Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - An intracellular stain was performed on RH30 cells with TLR4 Antibody (76B357.1) NB100-56566 (blue) and a matched mouse IgG2b Kappa isotype control (orange) MAB004. Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Human skin stained with at 5 ug/mL, peroxidase-conjugate and DAB chromogen. Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min.

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis of TLR4 in FFPE human colon tissue using an isotype control (top) and NB100-56566 (bottom) at 5 ug/mL.

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Human testis tissue stained with antibody at 5 ug/mL.

Immunohistochemistry-Frozen: TLR4 Antibody (76B357.1) [NB100-56566] - This image is TLR4(green) and nucleus(blue) at area postrema of the adult male mouse brain, x20 magnification. Primary antibody diluted 1:500. IHC-Fr image submitted by a verified customer review.

Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis of formaldehyde fixed THP-1 cells (human monocytic leukemia cells) using 2 ug/10^6 cells TLR4 antibody (clone 76B357.1) with detection employing a donkey anti-mouse IgG (H+L) cross adsorbed secondary antibody, (DyLight 488 conjugated). Isotype control samples incubated with mouse IgG2b isotype control antibody were processed in parallal under the same assay conditions.

Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - An intracellular stain was performed on Jurkat cells with TLR4 Antibody (76B357.1) NB100-56566 (blue) and a matched mouse IgG2b Kappa isotype control (orange) MAB004. Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).

Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - An intracellular stain was performed on Raw264.7 cells with TLR4 Antibody (76B357.1) NB100-56566 (blue) and a matched mouse IgG2b Kappa isotype control (orange) MAB004. Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).

Representative images of immunohistochemical staining of pancreatic adenocarcinoma upregulated factor (PAUF) and toll-like receptor 4 (TLR4) in human ovarian cancer specimens. The top row (A,B) represents normal ovarian tissue, middle row (C,D) shows immunoreactivity of ovarian carcinomas and the bottom row (E,F) indicates immunonegativity of ovarian caricinoma. The stromal cells of normal ovarian tissue show immunonegativity for PAUF and TLR4. The scale bar represents 100 μm.

ALWPs significantly reduced LPS-induced cell-surface levels of TLR4. (C) BV2 microglial cells were pretreated with ALWPs (500 ug/ml) or PBS for 30 min, followed by treatment with LPS (1 ug/ml) or PBS for 5.5 h. Cell-surface biotinylation was then conducted with a TLR4 antibody recognizing the N-terminal region of TLR4. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30319390), licensed under a CC-BY licence.

Effect of Cucumis sativus L. extract (CSE) on LPS induced TLR4 expression. b) Representative Western Blot of TLR4 and relative housekeeping alpha-tubulin were reported. c Expression of TLR4 protein in pAECs treated with LPS (10 ug/ml) in the presence or absence of increasing doses CSE (0.02; 0.2; 2 mg/ml); data shown are representative of at least three independent experiments and represent the mean +/- SEM. Different letters above the bars indicate significant differences (p < 0.05 ANOVA post hoc Tukey's test) Image collected and cropped by CiteAb from the following publication (//bmccomplementalternmed.biomedcentral.com/articles/10.1186/s12906-018-2254-1), licensed under a CC-BY licence.

Dasatinib affects TLR4 signaling to regulate LPS-induced proinflammatory cytokine levels in BV2 microglial cells. d) BV2 microglial cells were treated with dasatinib (250 nM) or vehicle (1% DMSO) for 30 min followed by LPS (1 ug/ml) or PBS for 5.5 h, and cell surface biotinylation was conducted. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31655606), licensed under a CC-BY licence.

Corroboration of human AKR2–protein interactions with highest confidence. (A) Representation of the components of the protein pull-down experiment using c-Myc magnetic beads with Myc-tagged human AKR2. (B) Western blot analysis of AKR2–protein interactions. Interacting proteins were incubated with AKR2 and immunopresipitated with c-Myc magnetic beads specific for the AKR2 protein tag (I) or with the c-Myc magnetic beads only as negative control (C-). Recombinant interacting proteins were included as positive control (C+). Mouse or rabbit antibodies specific for each protein were used as primary antibodies and then anti-mouse or anti-rabbit secondary antibodies were used to identify the presence of the interacting proteins (red arrow). The origin of the primary antibody is shown. (C) Corroboration of human AKR2–AKR2 interaction using protein pull-down with c-Myc magnetic beads with human Myc-tagged AKR2. The experiment was conducted as described in (B). The origin of the primary antibody and predicted size for AKR2 monomer, dimer, and trimer together with bands corresponding to fragments of the mouse anti-c-Myc antibody attached to magnetic beads are shown. (D) In order to identify unrelated proteins eluted from the c-Myc magnetic beads, a similar experiment was conducted with selected proteins but incubating only with the secondary antibody. A lane of SDS-PAGE stained with Bio-Safe Coomassie Stain corresponding to c-Myc magnetic beads alone eluted using Laemmli sample buffer was included (C--). The bands corresponding to fragments of the mouse anti-c-Myc antibody attached to magnetic beads are shown. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32046307), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$TDP1 depletion sensitizes glioblastoma-resistant cancer cells to temozolomide. (a) Cell lysates from RKO and DLD1 cancer cell lines were fractionated by SDS–PAGE and analyzed by immunoblotting using anti-TDP1 (Eurogentec), anti-Top1 (Santa Cruz), anti-Tubulin (Abcam) and anti-APE1 (Novus) antibodies (left). RKO and DLD1 cells were compared for their survival following exposure with the indicated doses of MMS for 15 min at 37°C (right). Data are the average of three independent experiments ± s.e.m. Where not visible, error bars are smaller than the symbol. (b) RKO cells were subjected to scrambled siRNA (Mock) or siRNA against TDP1 (TDP1KD) and cell lysates analyzed by immunoblotting (left). Control RKO (RKOSc siRNA) and RKO cells in which TDP1 levels were depleted (RKOTDP1 siRNA) were examined for their survival following exposure to the indicated doses of MMS, as described earlier in the text (right). (c) Glioblastoma multiforme T98G and U87 cancer cell lines were analyzed for methylguanine methyltransferase (MGMT) expression using anti-MGMT antibodies (Abcam) (left). High-MGMT expressing T98G cells were compared with low-MGMT expressing U87 cells for their survival following exposure to increasing concentrations of the alkylating agent temozolomide (right). (d) TG98 cells were subjected to scrambled siRNA (Mock) or siRNA against TDP1 (TDP1KD) and cell lysates analyzed by immunoblotting (left). Control T98G cells (T98GSc siRNA) and cells in which TDP1 levels were depleted (TG98TDP1 siRNA) were incubated with DMSO or 150 μM of the APE1 inhibitor CRT0044876 ‘APEi’ for 2 h followed by additional incubation with the indicated concentrations of MMS for 15 min at 37°C. Cell survival was blindly scored from three independent biological repeats, and data are the average ± s.e.m. (right). (e) Model for the repair of temozolomide-induced DNA breaks. Temozolomide delivers a methyl group to purine bases of DNA, resulting in O6-methyl guanine, N7-methyl guanine and N3-methyl adenine. The primary cytotoxic lesion is believed to be O6-methyl guanine, which can be removed by a direct repair mechanism mediated by methylguanine methyltransferase (MGMT). Inherent and acquired resistance to temozolomide via MGMT expression presents a major challenge in cancer therapy, particularly for glioblastoma multiforme. TDP1 promotes the repair of methylated purines induced by temozolomide via a distinct non-canonical BER pathway. Increasing the load of unrepaired methylated purines by exploiting the limited availability of TDP1 alone or in combination with canonical BER factors such as APE1 provides a new synthetic lethal setting to improve the clinical outcome of temozolomide-based cancer therapy. Asterisks denote statistical differences (P < 0.05; t-test) between control and TDP1-deficient cells. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/24335147), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

OTULIN overexpression reduced the number of Iba-1+ microglial cells in the cortical ischemic penumbra in focal cerebral ischemia/reperfusion rats. A Representative images of Iba-1+ microglial cells that were morphologically classified into three types; scale bar = 10 μm. B Iba-1+ microglial cells in the ischemic penumbra and contralateral cortex of each group were detected by confocal immunofluorescence at 24 h, 72 h, and 7 days; scale bar = 75 μm. C The histogram represents the quantitative analysis of mean Iba-1 immunofluorescence intensity in each group (n = 6). Data are presented as the means ± SEMs; ###P < 0.001 versus the tMCAO group; &&&P < 0.001 versus the tMCAO+LV-Scramble group Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29544517), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

FKBP51 modulates EMT hallmarks. (a and b) FKBP51 silencing of SAN melanoma cells decreases TGF-beta and T beta RIII expression. Normalized expression rates (mean±S.D.) of TGF-beta (a, left) and T beta RIII (b, left) mRNA levels; (a, n=3; b, n=5). FKBP51 siRNA-treated sample expression=1. Western blot assay of TGF-beta in SAN melanoma cells treated with nonsilencing (NS) RNA or FKBP51 siRNA (a, right). Flow cytometric analysis of T beta RIII (endoglin) expression (b, right): FKBP51 siRNA-treated cells moved to the left relative to NS RNA-treated cells because of reduced staining (MFI: 111±10 and 64±3 in NS RNA and FKBP51 siRNA-treated cells, respectively). (c, and d) FKBP51 overexpression enhances TGF-beta and T beta RIII expression. Normalized expression of TGF-beta (c, left) and T beta RIII (d, left) mRNA in WT, EV-, or FKBP51-stably transfected melanoma cells. WT sample expression=1. (c, n=3; d, n=5). Western blot assay of TGF-beta levels in the same cells (c, right). Anti-Flag-labelled exogenous FKBP51. Flow cytometric analysis of T beta RIII expression (d, right). FKBP51-overexpressing cells moved to the right relative to WT or EV cells, indicating increased expression. (e) Graphic representation of cell counts ( × 10−3) in WT, EV-, or FKBP51-stably transfected melanoma cell cultures. Results are means of six independent experiments each performed in triplicate. (f) FKBP51 upregulates colony number and size. Colonies were counted under low magnification ( × 10) at four points on each dish. The data represent the means of two independent experiments performed in triplicate. Magnification: upper, × 10; lower × 40. (g) Boyden chamber filters invaded by melanoma cells. Filter was stained with Crystal Violet. OD value was obtained by reading the eluted Crystal Violet with a spectrophotometer at a wavelength of 570 nm. The increased OD in FKBP51 samples indicated that FKBP51-overexpressing cells invaded matrigel more efficiently than control cells. The data represent the means of three independent experiments performed in triplicate Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/cddis2013109), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Functional assessment of Fanconi anemia pathway. (a) The experimental scheme for MMC treatment. Twenty‐four hours after plating, cells were cultured with or without MMC 1 μM for an additional 24 hr, after which the cells were harvested for western blot or immunostaining. (b) Western blot with FANCD2 antibody of BJ, proband fibroblasts, FANCB mutant (null) fibroblasts and FANCD2 mutant (null) fibroblasts. (c) Western blot with FANCD2 antibody of non‐FA control lymphoblasts (LCL), proband LCL‐A‐2017, proband LCL‐B‐2017, FANCB mutant (null) LCL, and FANCD2 mutant (null) LCL. (d) Representative figures of FANCD2 foci formation in the indicated cells. (e) Quantification of FANCD2 foci formation following treatment with or without 1 μM MMC. Experiments were performed in triplicate. One hundred cells were counted for each experiment Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29193904), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Pot1a prevents DDR in HSCs. a, b Telomeric DDR in 8 week-old LSK cells upon Pot1a knockdown. a Immunocytochemical staining of TRF1 (green) and 53BP1 (red). Foci co-stained with TRF1 and 53BP1 were identified as TIFs. Nuclei were stained with TOTO3 (blue). Scale bar, 2 μm. b Frequency of TIFs per cell after 1 week of culture. Data are expressed as the mean ± SD (n = 100, *p < 0.01 by t-test). Representative data from three independent experiments are shown. c–f Telomeric DDR in donor-derived control-GFP or Pot1a overexpressing 8 week-old LSK cells 4 months post BMT. c Immunocytochemical staining of TRF1 (green) and 53BP1 (red). Nuclei were stained with TOTO3 (blue). Scale bar, 2 μm. d Frequency of TIFs per cell. Data are expressed as the mean ± SD (n = 120, *p < 0.01 by t-test). Representative data from 3 independent experiments are shown. e Immunocytochemical staining of TRF1 (green) and RPA32 (red). f Immunocytochemical staining of TRF1 (green) and pChk1 (red). Scale bar, 2 μm. g Flow cytometric analysis of Chk1 and pChk1 in donor-derived GFP+ LSKCD48–CD150+ cells after 5 months of 2nd BMT. Mean fluorescence intensity of Chk1 and pChk1 (left panels). Data are expressed as the mean ± SD (n = 3, *p < 0.01 by t-test). Representative FACS profiles of Chk1 and pChk1 in donor-derived GFP+ LSKCD48–CD150+ cells (right panels) Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28986560), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Lamin A/C-53BP1 interaction is regulated in a DNA damage-dependent manner. (A) Left; HDF were subjected to IR (10 Gy) and allowed to recover for 1 h. Association between A-type lamins and 53BP1 was assessed by immunoprecipitation of endogenous lamin A/C followed by immunoblotting with 53BP1 and lamin A/C antibodies. WCE, whole cell extract, IP: immunoprecipitates. WCE represents 5% input. Right; endogenous lamin A/C was immunoprecipitated and supernatants or pellets were analysed by immunoblotting for interaction with JMJD2A. (B) U2OS/GFP-lamin A cells were pretreated with caffeine (20 mm) for 1 h before exposure to IR (10 Gy, 1 h recovery). Cell extracts were then subjected to immunoprecipitation using GFP-Trap beads, and bound complexes were then analysed by immunoblotting using 53BP1 and GFP antibodies. WCE represents 1% input. (C) As in (C) except cells were pretreated with 10 μm ATMi for 1 h before IR. WCE represents 1% input. (D) U2OS/GFP-lamin A cells were subjected to laser micro-irradiation, fixed 1 h later and immunostained with gamma -H2AX antibody. Scale bar, 10 μm. (E) U2OS cells were transfected with siCTRL or siLMNA and subjected to laser micro-irradiation, fixed 15 min later and then processed for immunofluorescence with gamma -H2AX and 53BP1 antibodies. Scale bar, 10 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/25645366), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Bv, Ma, Pm, ChHa

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Ma, Pm, ChHa

Applications:

, , , , , , , , , , , , , ,

Applications: , , , , , , , , , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Alt Catalog#

IMG-5031A

Alt Catalog# IMG-5031A

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP2-27149AF350
Alexa Fluor 405	NBP2-27149AF405
Alexa Fluor 488	NBP2-27149AF488
Alexa Fluor 532	NBP2-27149AF532
Alexa Fluor 594	NBP2-27149AF594
Alexa Fluor 647	NBP2-27149AF647
Alexa Fluor 700	NBP2-27149AF700
Alexa Fluor 750	NBP2-27149AF750
Allophycocyanin	NBP2-27149APC
Allophycocyanin/Cy7	NB100-56566APCCY7
Biotin	NBP2-27149B
DyLight 350	NBP2-27149UV
DyLight 405	NBP2-27149V
DyLight 488	NBP2-27149G
DyLight 550	NBP2-27149R
DyLight 594	NB100-56566DL594
DyLight 650	NBP2-27149C
DyLight 680	NBP2-27149FR
DyLight 755	NBP2-27149IR
FITC	NBP2-27149F
HRP	NBP2-27149H
Janelia Fluor 549	NB100-56566JF549
Janelia Fluor 646	NB100-56566JF646
PE	NBP2-27149PE
PE/Atto594	NB100-56566PEATT594
PE/Cy5.5	NB100-56566PECY55
PE/Cy7	NB100-56566PECY7
PerCP	NBP2-27149PCP
mFluor Violet 450 SE	NBP2-27149MFV450
mFluor Violet 500 SE	NBP2-27149MFV500
mFluor Violet 610 SE	NBP2-27149MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Azide Free	NBP2-27149
BSA Free	NB100-56566SS
Azide Free	NBP2-27149SS

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AF3626

IL-33 Antibody [Unconjugated]

(2 Reviews)

(139 Publications)

100 μg

AF3626

Western blot shows lysates of HEK293T human embryonic kidney cell line either mock transfected or transfected with full length mouse IL-33. PVDF membrane was probed with 0.4 µg/mL of Goat Anti-Mouse IL‑33 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3626) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for IL‑33 at approximately 37 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

IL‑33 was detected in immersion fixed frozen sections of mouse spleen using Goat Anti-Mouse IL‑33 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3626) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

IL‑33 was detected in immersion fixed bEnd.3 mouse endothelioma cell line using Goat Anti-MouseIL‑33 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3626) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red, upper panel; Catalog # NL001) and counterstained with DAPI (blue, lower panel). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Mouse splenocytes were treated for 5 hours with 50 ng/mL PMA and 1 μg/mL Ca2+ ionomycin then stained with Goat Anti-Mouse IL‑33 Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF3626, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Recombinant Mouse IL‑33 (Catalog #3626-ML) stimulates proliferation in the D10.G4.1 mouse helper T cell line in a dose-dependent manner (orange line), as measured by Resazurin (Catalog # AR002). Proliferation elicited by Recombinant Mouse IL‑33 (0.25 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse IL‑33 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3626). The ND50 is typically 10-50 ng/mL.

IL-33/ST2 axis in tendon healing in vivo.Kinetics of (a) Il33 and (b) soluble St2 gene expression post injury. Data are mean fold change±s.d. showing relative expression to 18s housekeeping gene (pooled data from four mice per group performed on four sequential occasions, n=16 per condition). *P<0.05, **P<0.01 versus control (WT) mice (ANOVA) +P<0.05,++P<0.01, NS not significant; control (WT) injured versus St2−/− injured. (c) Immunohistochemistry showing IL-33 and ST2 expression in tendon biopsies of WT and St2−/− mice day 1 post injury. IgG control shown in bottom left of pictures. Black horizontal line indicates 50 μm. (d) Col3 mRNA and (e) Collagen 3 protein levels in tendon biopsies of WT and St2−/− mice post injury. (f) Col1 mRNA and (g) Collagen 1 protein levels in tendon biopsies of WT and St2−/− mice post injury. For (d–g) data are mean±s.d. of duplicate samples, representative of four mice per condition (n=16). *P<0.05, **P<0.01 versus control (WT) mice. +P<0.05, ++P<0.01 control (WT) injured versus St2−/− injured mice (ANOVA). mRNA graphs show relative expression to 18s housekeeping gene. (h) Percentage change in tendon strength for WT and St2−/− mice post injury. Data are mean±s.d., representative of four mice per condition (n=16). *P<0.05, **P<0.01 versus control mice. #P<0.05 St2−/− injured versus WT injured mice (Mann–Whitney U-test). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25857925), licensed under a CC-BY license. Not internally tested by R&D Systems.

HMGB1 increases IL-33 expression and acts as an autocrine factor in enhancing IL-33 expression in a partial TLR-4 dependent fashion during cyclic stretch.(A) MLE-12 cells were treated with 3 μg/ml HMGB1 or control solution before 6h cyclic stretch and IL-33 production was measured by ELISA. (B) MLE-12 cells were treated with 10 μg/ml HMGB1 neutralizing antibody (2G7) or control solution before 6h cyclic stretch and IL-33 production was detected by Western blot. beta -actin served as loading control. (C) MLE-12 cells transfecting with non-specific control siRNA or TLR-4-specific siRNA were treated with 3 μg/ml HMGB1 or control solution before stretch. IL-33 production in each group was measured with ELISA. **P<0.01, ***P<0.001 when compared between groups denoted by horizontal lines. Each stretching group collected at least from three wells and represented a single experiment, the bar graphs illustrate data representative of three independent experiments. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28898270), licensed under a CC-BY license. Not internally tested by R&D Systems.

IL-33/ST2 expression in human tendon.(a) IL-33, (b) soluble ST2 (sST2) and (c) mST2 mRNA expression in human tendon samples. Fold change in gene expression of IL-33, sST2 and mST2 in control (semi-membranosus tendon, n=10), torn supraspinatus tendon (established pathology) and matched subscapularis human tendon samples (early pathology; n=17). Data are mean±s.d. relative to the housekeeping gene18S (mean of duplicate analysis). *P<0.05, **P<0.01, ***P<0.001 versus control (Student's t-test). (d) Immunostaining of IL-33 and ST2 in control (n=10), torn tendon (n=17) and early tendinopathy (n=17). Graphs illustrate modified Bonar scoring based on 10 high-power fields. Data are mean±s.d. *P<0.05, **P<0.01 versus control (Student's t-test). Scale bar, 65 μm. (e) Fold change in gene expression of IL33 and ST2 24 h post incubation with tumour necrosis factor (TNF-alpha ), IL-1 beta or in combination depicting relative expression to media alone utilizing housekeeping gene GAPDH. Data are mean±s.d. of triplicate samples, representative of three individual patient samples. *P<0.05, **P<0.01 versus control (media) (Student's t-test). (f) IL-33 immunostaining of human tendon explants cultured for 24 h with medium (control), 100 ng ml−1 TNF alpha or 100 ng ml−1 TNF alpha +100 ng ml−1 IL-1 beta . (g) Fold change in COL1 and COL3 mRNA expression in human tendon explants cultured for 24 h with rhIL-33, relative to housekeeping gene GAPDH. (h) Time course of COL1 and COL3 mRNA expression following incubation with rhIL-33, relative to housekeeping gene GAPDH. (i) Collagen 1 and 3 protein expression in human tendon explants 24 h post incubation with rhIL-33. For (g–i) data are mean±s.d. of triplicate samples, representative of three individual patients. *P<0.05, **P<0.01 versus control (Student's t-test). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25857925), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Biotin	BAF3626

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF3626-SP

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NB600-503

beta-Actin Antibody

(4 Reviews)

(123 Publications)

0.05 ml

NB600-503

Simple Western: beta-Actin Antibody [NB600-503] - Simple Western lane view shows a specific band for Beta Actin in 0.1 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Flow Cytometry: beta-Actin Antibody [NB600-503] - An intracellular stain was performed on HeLa cells with beta-actin Antibody NB600-503AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

Western Blot: beta-Actin Antibody [NB600-503] - Western blot analysis of Jurkat, COS, and CHO cell lysate using beta actin antibody at 1:100.

Immunohistochemistry-Paraffin: beta-Actin Antibody [NB600-503] - Analysis of Beta Actin in mouse epidermis using DAB with hematoxylin counterstain.

Immunocytochemistry/Immunofluorescence: beta-Actin Antibody [NB600-503] - IF Confocal analysis of HeLa cells using Beta Actin antibody (NB600-503, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). DAPI was used to stain the cell nuclei (blue).

Flow Cytometry: beta-Actin Antibody [NB600-503] - Analysis of HeLa cells using mouse Monoclonal beta-Actin antibody (Orange) and Isotype control Antibody (Blue).

Simple Western: beta-Actin Antibody [NB600-503] - Early start GM1 administration did not affect alpha-synuclein expression or transport to the striatum. When assessed 1 week following AAV-A53T alpha-synuclein injection, levels of striatal alpha-synuclein were no different in saline (N=6) vs. GM1-treated animals (N=6), suggesting no influence of GM1 on A53T alpha -synuclein transduction or transport to the striatum. Representative Wes Simple Western blots are shown after cropping (full length images of blots are presented as Supplementary Fig. 2. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-019-42847-x), licensed under a CC-BY license.

Early start GM1 administration did not affect alpha-synuclein expression or transport to the striatum. (A,B) When assessed 1 week following AAV-A53T alpha-synuclein injection, levels of striatal alpha-synuclein were no different in saline (N = 6) vs. GM1-treated animals (N = 6), suggesting no influence of GM1 on A53T alpha -synuclein transduction or transport to the striatum. Representative Wes Western blots are shown after cropping (full length images of blots are presented as Supplementary Fig. 2. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31182727), licensed under a CC-BY licence.

Anti-hnRNP A1 antibodies alter protein levels as measured by Western blot. SK-N-SH cells were cultured and treated with anti-hnRNP A1 antibodies or control IgG. Following a 48 hour incubation, cells were lysed and protein lysate was run on 10% Tris-glycine gels for Western blot analysis and probed for Beta-actin (A) or GAPDH (B)) (control), hnRNP A1, SPG4, SPG7, and SPG20. Results revealed that there was a marked reduction of SPG 4 and SPG 7 protein levels in anti-hnRNP A1 antibody compared to control isotype IgG treated cells. There was a variable response to hnRNP A1 protein and a modest reduction of SPG 20. Parentheses show relative percent reduction of signal comparing anti-hnRNP A1 antibody to control isotype IgG treatment of cells. (L=lysate, IgG=control isotype IgG, A1=anti-hnRNP A1-M9 antibody treatment of cells). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27375925), licensed under a CC-BY licence.

SPOP expression in gastric cancer. (A) The representative photos of SPOP expression in gastric cancer tissues and adjacent gastric tissues by using immunohistochemical staining (DAB staining, scale bar, 40 μm). Magnified local images reflecting detailed information were shown on the bottom. (B, C) SPOP expression was plotted using the immunochemical scores as described in the Methods. B, plot of SPOP scores in each gastric carcinoma and adjacent tissues. C, scores of SPOP expression are shown as box plots. The horizontal lines represent the median. The bottom and top edges of the boxes represent the 25th and 75th percentiles, respectively. And the vertical bars represent the range of the data. n = 88. (D) Representative immunoblotting of SPOP protein in gastric cancer samples. C refers to gastric cancer tissue and A refers to paired adjacent non-tumor gastric tissue from the same patient. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/25204354), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Av, Bv, ChHa, Fi, Pm, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Av, Bv, ChHa, Fi, Pm, Rb

Applications:

, , , , , , , ,

Applications: , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 405	NB600-503AF405
Alexa Fluor 488	NB600-503AF488
Alexa Fluor 647	NB600-503AF647
Alexa Fluor 700	NB600-503AF700
Allophycocyanin	NB600-503APC-0.05ml
Biotin	NB600-503B
DyLight 350	NB600-503UV-0.05ml
DyLight 405	NB600-503V
DyLight 488	NB600-503G
DyLight 550	NB600-503R
DyLight 594	NB600-503DL594
DyLight 650	NB600-503C
DyLight 680	NB600-503FR-0.05ml
DyLight 755	NB600-503IR-0.05ml
FITC	NB600-503F
HRP	NB600-503H
Janelia Fluor 549	NB600-503JF549
Janelia Fluor 646	NB600-503JF646
PE	NB600-503PE-0.05ml
PerCP	NB600-503PCP-0.05ml

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At	=	A. thaliana	SyHa	=	Golden Syrian Hamster
All	=	All Species	Ha	=	Hamster
An	=	Animal	Hu	=	Human
ArHa	=	Armenian Hamster	I	=	Insect
Av	=	Avian	Ll	=	Llama
Bb	=	Baboon	Ma	=	Mammal
Ba	=	Bacteria	Mk	=	Monkey
Bv	=	Bovine	Mu	=	Mouse
Ca	=	Canine	Multi	=	Multi-species
Ce	=	C. Elegans	NA	=	Non-species specific
Ch	=	Chicken	Or	=	Orangutan
ChHa	=	Chinese Hamster	Pl	=	Plant
Cm	=	Cynomolgus Monkey	Pm	=	Primate
Do	=	Donkey	Po	=	Porcine
Dr	=	Drosophilia	Rb	=	Rabbit
Ec	=	E. Coli	Rt	=	Rat
Eq	=	Equine	RM	=	Rhesus Macaque
Fe	=	Feline	Sh	=	Sheep
Ft	=	Ferret	Vb	=	Vertebrate
Fi	=	Fish	Vi	=	Virus
Fu	=	Fungi	Xp	=	Xenopus
Ge	=	Gerbil	Ye	=	Yeast
GP	=	Guinea Pig	Ze	=	Zebrafish
Gt	=	Goat

Category

Protein/Target

Applications

Reactivity

Citation

Antibody Validation

Reviews

Host

Available Conjugates

Format

Marker

Modification

Clonality

Clone

Gene Symbol

Images Type

Inhibitor Family

Inhibitor Target

Isotype

Preservative

Protein Or Peptide

Research Areas

Has Sample Size

Promotional Item

Species Glossary

IL-1 beta/IL-1F2 Antibody [Unconjugated]

ABCA1 Antibody - BSA Free

ACE-2 Antibody [Unconjugated]

PDGFR alpha Antibody [Unconjugated]

SR-BI Antibody - BSA Free

TGF beta 1/2/3 Antibody (1D11) [Unconjugated]

Osteopontin/OPN Antibody [Unconjugated]

CXCL8/IL-8 Antibody (6217) [Unconjugated]

TLR9 Antibody (26C593.2) - BSA Free

TLR4 Antibody (76B357.1) - BSA Free

IL-33 Antibody [Unconjugated]

beta-Actin Antibody