IL-1 beta/IL-1F2 Antibodies: Novus Biologicals

AF-401-NA

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

(3 Reviews)

(517 Publications)

100 μg

AF-401-NA

Western blot shows lysates of THP-1 human acute monocytic leukemia cell line untreated (-) or treated (+) with 200 nM PMA for 24 hours and 10 µg/mL LPS for 4 hours and RAW 264.7 mouse monocyte/macrophage cell line untreated (-) or treated (+) with 10 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 (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; 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; 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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NB600-633

IL-1 beta/IL-1F2 Antibody

(4 Reviews)

(92 Publications)

0.1 mg

NB600-633

Immunohistochemistry: IL-1 beta/IL-1F2 Antibody [NB600-633] - Analysis of: Human IL1beta antibody Secondary antibody: Peroxidase goat anti-rabbit at 1:10,000 for 45 min at RT Localization: cytoplasm Staining: Close up of medullary lymph node: positive staining in the cytoplasm of circulating macrophages. Neg Ctr (far right) normal rabbit IgG with pH 6.2 40X

Western Blot: IL-1 beta/IL-1F2 Antibody [NB600-633] - Analysis of IL-1 beta in Rhesus fetal membrane using anti-IL-1 beta antibody. Image from verified customer review.

Western Blot: IL-1 beta/IL-1F2 Antibody [NB600-633] - Antibody was used at a 1:200 dilution incubated 1h at room temperature to detect dog IL-1b by Western blot. Lane 1 and 2 were loaded with 2.5 ug and 500 ng of dog IL-1b respectively. The molecular weight of the detected band is estimated by comparison to molecular weight markers (not shown). Detection occurred using a 1:3,000 dilution of IRDye (TM) 800 conjugated Donkey anti-Rabbit IgG for 1h at room temperature.

Western Blot: IL-1 beta/IL-1F2 Antibody [NB600-633] - pro-IL-1beta in glioblastoma cells. Image from verified customer review.

Western Blot: IL-1 beta/IL-1F2 Antibody [NB600-633] - Lane 1: Human IL-1Beta. Load: 5 ng per lane. Primary antibody: Human IL-1Beta antibody at 1:2,000 for overnight at 4C. Secondary antibody: Peroxidase rabbit secondary antibody at 1:40,000 for 30 min at RT. Block: Blocking Buffer for Fluorescent Western Blotting for 30 min at RT. Predicted/Observed size: 17 kDa, 17 kDa for Human IL-1Beta. Other band(s): Unspecific band at 35 kDa.

Western Blot of Rabbit anti-Human IL-1beta antibody. Lane 1: Human IL-1beta. Load: 5 ng per lane. Primary antibody: Human IL-1beta antibody at 1:2,000 for overnight at 4C. Secondary antibody: Peroxidase rabbit secondary antibody at 1:40,000 for 30 min at RT. Block: Blocking Buffer for Fluorescent Western Blotting () for 30 min at RT. Predicted/Observed size: 17 kDa, 17 kDa for Human IL-1beta. Other band(s): Unspecific band at ~35 kDa.

$miR-27a/b prevent Parkin translocation to mitochondria upon mitochondrial damage. a, b 48 h post-transfection, HeLa cells were incubated with 10 μM CCCP for 2 h. After isolating the mitochondria-enriched fractions, Parkin translocation and LC3-II were assessed by Western blot. Each protein level was normalized to corresponding mitochondrial loading control (SOD2) level and quantified as a percentage of control (n = 3, two-way ANOVA). Purity of mitochondrial fraction was assessed by monitoring p38 cytosolic and VDAC1 mitochondrial markers. PN; post-nuclear, C; cytoplasm, M; mitochondrial (A). c, d 48 h post-transfection, HeLa cells stably expressing GFP-Parkin were visualized by GFP, a mitochondrial marker (TOM20) antibody, and a nuclear dye (Hoechst) as indicated. Scale bars correspond to 10 μm. Parkin translocation was quantified as the ratio of cytoplasmic to nuclear GFP signal and the resulting ratios were normalized to control. Data were collected from 4 independent replicates (n > 1000 cells) and are shown as a percentage of control. Values are mean ± SEM (*p < 0.05, **p < 0.01, ***p < 0.001) Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27456084), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Expression of PACAP, VIP, VIPRs, HIF-1 alpha , HIF-2 alpha , and EGFR in glioblastoma multiforme (GBM). (A) Representative immunoblot of PACAP and VIP precursor peptides and PAC1R, VPAC1R, and VPAC2R expression on frozen glioblastoma sample. (B) Representative immunoblot and photomicrographs of signals detected by antibodies direct against HIF-1 alpha , HIF-2 alpha , and EGFR in a frozen glioblastoma sample. Image collected and cropped by CiteAb from the following open publication (//journal.frontiersin.org/Article/10.3389/fphar.2016.00139/abstract), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Ca, Pm, SyHa

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ca, Pm, SyHa

Applications:

, , , , , , , ,

Applications: , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Validated by:

Validated by:

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AF-201-NA

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

(3 Reviews)

(83 Publications)

100 μg

AF-201-NA

IL-1 beta /IL-1F2 mRNA was detected in formalin-fixed paraffin-embedded tissue sections of human tonsil probed with ACD RNAScope® Probe (Catalog # 310361) and stained using ACD RNAscope® 2.5 HD Detection Reagents-Red (top image, Catalog # 32260). Adjacent tissue section was processed for immunohistochemistry using R&D Systems Goat Anti-Human IL‑1 beta /IL‑1F2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-201-NA) at 1 ug/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte HRP Polymer Antibody (R&D Systems, Catalog # VC004) and DAB chromogen (lower image, yellow-brown). Tissues were counterstained with hematoxylin (blue).

<b>Orthogonal Strategies Validation. </b>Western blot shows lysates of THP-1 human acute monocytic leukemia cell line untreated (-) or treated (+) with 200 nM PMA for 24 hours and 10 µg/mL LPS and 3 hours. PVDF membrane was probed with 0.1 µg/mL of Goat Anti-Human IL-1 beta /IL-1F2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-201-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 36 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 human peripheral blood mononuclear cells (PBMCs) treated with LPS and monensin using Goat Anti-Human IL-1 beta /IL-1F2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-201-NA) at 10 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). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

IL‑1 beta /IL‑1F2 was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human IL‑1 beta /IL‑1F2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-201-NA) at 1 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Simple Western lane view 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 hrs and 3 hrs, respectively, and loaded at 0.2 mg/mL. A specific band was detected for IL‑1 beta /IL‑1F2 at approximately 39 kDa (as indicated) using 1 µg/mL of Goat Anti-Human IL‑1 beta /IL‑1F2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-201-NA) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Recombinant Human IL‑1 beta /IL‑1F2 (Catalog # 201-LB) stimulates proliferation in the the D10.G4.1 mouse helper T cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human IL‑1 beta /IL‑1F2 (50 pg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human IL‑1 beta /IL‑1F2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-201-NA). The ND50 is typically 0.004-0.02 µg/mL.

BTK inhibitors and its dysfunctional mutation suppress NLRP3 inflammasome activation.(a) Enzyme-linked immunosorbent assay (ELISA) of human IL-1 beta in supernatants and immunoblot analysis of human IL-1 beta p17, caspase-1 p20/p22 in supernatants and pro-IL-1 beta in cell lysates of THP-1-Mφs that were pretreated with the indicated inhibitors for 30 min and then stimulated with alum for 6 h. ELISA of murine IL-1 beta (b,c) and TNF-alpha (b) in supernatants of LPS-primed murine peritoneal macrophages that were pretreated with LFM-A13 and then stimulated with alum for 3 h. (d,e) Immunoblot analysis of the indicated proteins (d) and ELISA of murine IL-1 beta and IL-6 in supernatants of LPS-primed peritoneal macrophages from Xid and WT mice stimulated with alum for 6 h. (f) ELISA of murine IL-1 beta in supernatants of LPS-primed murine peritoneal macrophages pretreated with LFM-A13 and/or Syk inhibitor (R406), then stimulated with alum for 3 h. (g) ELISA of murine IL-1 beta and immunoblot analysis of murine caspase-1 p20 in supernatant of LPS-primed murine peritoneal macrophages stimulated with the indicated NLRP3 inflammasome activators for 3 h. Immunoblot analysis of murine IL-1 beta p17, caspase-1 p20 in supernatants, pro-IL-1 beta , pro-caspase-1 and ASC in cell lysates (h), and ELISA of murine IL-1 beta in supernatants (i) of LPS-primed murine peritoneal macrophages that were pretreated with LFM-A13 and then stimulated with alum or poly(dA:dT) for 3 h. Data are representative of three independent experiments. Data are presented as mean±s.d. (triplicate). **P<0.01; ***P<0.003. Two-sided Student's t-test. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26059659), licensed under a CC-BY license. Not internally tested by R&D Systems.

Pyroptosis/IL-1 beta pathway is enhanced in vivo and can be modulated by anakinra. a In the PI model, phosphorylated-p65 NF-kappa B, NLRP3, 8-OHdG, IL-1 beta , and TUNEL were detected by IHC (Bar, 100 μm). b IL-1 beta was immuno-localized in gum tissues of mice with PI alone and with orthotopic mammary tumor introduction (Bar, 100 μm). c The mRNA expression of IL-1 beta was measured in the gingiva of the indicated groups. Serum IL-1 beta was detected by ELISA (n ≥ 5; Student’s t test). d The size of primary tumors was measured (n ≥ 7). e Early metastasis in lymph nodes were measured by luciferase assay and normalized by protein concentration, where anakinra reduced the metastasis to the axillary and cervical lymph nodes. MDSC content was also decreased by anakinra in cervical lymph node. (n ≥ 7; Ank anakinra; ANOVA, *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001) Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31685946), licensed under a CC-BY license. Not internally tested by R&D Systems.

TMZ inhibits tumor growth and enhances the expression of NLRP1, IL-1 beta , and Notch1 in 1205Lu tumors in vivo. (A) Tumor growth curve of 1205Lu parental cells injected subcutaneously in mice, intraperitoneally treated with a TMZ cycle or 10% dimethyl sulfoxide (DMSO) (Control) (indicated by arrows). Tumor growth was monitored for 26 days. Data are expressed as the mean ± SEM. n = 10 tumors (control) or 12 tumors (TMZ). (B) qRT-PCR analysis of NLRP1 expression in single tumor cells isolated from the tumor tissues studied in (A). Data are expressed as the mean ± SEM. n = 3 samples. * p < 0.05 and *** p < 0.001 vs the corresponding tumors. (C–E) Immunohistochemical staining of tumor tissues with NLRP1 (C), IL-1 beta (D), and Notch1 (E). Samples MB656, MB657, and MB658 were from the control group, whereas MB659, MB660, and MB661 were from the TMZ group. Bar = 50 μm (left panels, 20×) or 20 μm (right panels, 63×). Representative images are shown. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32899791), licensed under a CC-BY license. Not internally tested by R&D Systems.

IL-1 and IL-1R1 expression is enriched in the melanoma stroma. (A) Real-time qPCR analysis of IL1A and IL1B expression in stage-III and stage-IV melanoma tumor samples (n = 39) relative to expression in human skin samples (n = 8). ***, P < 0.001; Mann-Whitney test. (B) Analysis of IL1B expression in normal skin and benign nevi samples (nonmalignant; n = 25) and cutaneous melanoma samples (malignant; n = 45) from an available gene expression dataset (Talantov et al., 2005) accessed through the Oncomine platform. (C) Sections from a case of primary cutaneous melanoma stained for IL-1 beta , CD163, and CD68 expression as indicated by labels. Bars: (i) 200 µm; (ii) 50 µm; (iii) 33 µm. (D) Serial sections from two skin metastases (i–iii and iv–vi, respectively), stained for IL-1R1 and SMA expression as indicated by the labels. Bars: (i and iv) 200 µm; (ii, iii, v, and vi) 33 µm. (C and D) Arrowheads indicate cells that are clearly double stained. (E) Western blot analysis of IL-1R1 and IL-1 beta precursor protein expression in a panel of cell lines. Data are representative of three independent experiments. (F) Secreted IL-1 beta in conditioned media from a panel of melanoma cell lines detected by ELISA. Data are represented as mean ± SEM for three independent samples in each group. **, P < 0.01; Dunn’s multiple comparisons test. (E and F) Macrophages (Mφ) were stimulated with 100 ng/ml IFN-gamma and 20 ng/ml LPS. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28450382), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

, , , , ,

Applications: , , , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Biotin	BAF201

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF-201-SP

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MAB601

IL-1 beta/IL-1F2 Antibody (2805) [Unconjugated]

(1 Review)

(63 Publications)

100 μg

MAB601

Western blot shows lysates of THP-1 human acute monocytic leukemia cell line untreated (-) or treated (+) with 200 nM PMA for 24 hours and 10 µg/mL LPS and 3 hours. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human IL-1 beta /IL-1F2 Monoclonal Antibody (Catalog # MAB601) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for IL-1 beta /IL-1F2 at approximately 36 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 human peripheral blood mononuclear cells (PBMCs) using Mouse Anti-Human IL-1 beta /IL-1F2 Monoclonal Antibody (Catalog # MAB601) at 8 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). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Effect of A1AT on whole blood IL-1 beta release. IL-1 beta production in whole blood cultures in response to LPS (1.0 μg/ml) was performed in the presence of endogenous A1AT (i.e., undiluted) or exogenously added A1AT (2 mg/ml) in blood diluted 1:32 with RPMI. Whole blood cultures were incubated for 18 h. After incubation, plasma supernatants were removed, and IL-1 beta quantified by ELISA and expressed as mean ± SD for three donors. The diluted sample result was corrected for the dilution. NS indicates no significant difference. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0117330), licensed under a CC-BY license. Not internally tested by R&D Systems.

Inflammatory response induced by HI injury and maternal SE. Representative images of immunofluorescence staining of inflammatory cytokines IL-1 beta , IL-6, and TNF alpha in the cerebral cortex (A) and the hippocampus (E). The immunofluorescence intensity of IL-1 beta (B,F), IL-6 (C,G), and TNF alpha (D,H) in the cerebral cortex and hippocampus. Results are presented as mean ± SEM. *P < 0.05, **P < 0.01, n = 4, analyzed by two-way ANOVA followed by post hoc Turkey tests. SH, from sham exposed dams with sham surgery; HI, hypoxic-ischemic injury; SE, from smoke exposed dams with sham surgery; HI + SE, from smoke exposed dams with hypoxic-ischemic injury. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35250486), licensed under a CC-BY license. Not internally tested by R&D Systems.

Effect of high glucose and IL-1 beta alone or in combination on the protein expressions of CD31, FSP1 and alpha -SMA in HAECs. a–d HAECs were incubated for 48 h with NG and HG. Mannitol was used as a control for hyperosmolarity. Representative western blots (a) and quantitative determinations of CD31, FSP1 and alpha -SMA protein levels (b–d) are presented. e–h HAECs were treated for 48 h with NG, HG, IL-1 beta (10 ng/ml)and HG in the presence of the IL-1 beta (10 ng/ml). Representative western blots (E) and quantitative determinations of CD31, FSP1 and alpha -SMA protein levels (f–h) are presented. The data are expressed as the mean ± SD. Experiments were repeated at least three times. NG normal glucose (5.5 mM), HG high glucose (30 mM), MN 5.5 mM glucose + 24.5 mM mannitol, IL-1 beta (10 ng/ml), HG + IL-1 beta : high glucose (30 mM) + IL-1 beta (10 ng/ml) *P < 0.05 vs. MN or NG, **P < 0.01 vs. NG, #P < 0.05 vs. HG Image collected and cropped by CiteAb from the following publication (//www.cardiab.com/content/15/1/42), licensed under a CC-BY license. Not internally tested by R&D Systems.

The influence of high glucose or IL-1 beta on immunofluorescence of CD31 and FSP1 in HAECs. Representative immunofluorescence images showing CD31 (green), FSP1 (red) labeling and DAPI (blue) stains nuclei. a Normal ECs monolayers displayed a cobble stone morphology. b A merge of the three images revealed some cells populations that acquired a spindle-shaped morphology and lost CD31 expression (white arrow). c HAECs exposure to IL-1 beta alone for 48 h acquired a spindle-shaped morphology. d High glucose and IL-1 beta in combination resulted in decreased CD31 (the left white arrow) and increased FSP1staining (the right arrow). a normal glucose (5.5 mM) group, b high glucose (30 mM) group for 48 h; c treatment with a normal glucose (5.5 mM) + IL-1 beta (10 ng/ml) treatment for 48 h, d treatment with a high glucose (30 mM) + IL-1 beta (10 ng/ml) treatment for 48 h. Scale bar, 75 μm Image collected and cropped by CiteAb from the following publication (//www.cardiab.com/content/15/1/42), licensed under a CC-BY license. Not internally tested by R&D Systems.

Effects of PKC beta on high glucose induced IL-1 beta up-regulation. Confluent cultures of HAECs were exposed to NG, HG, PMA (30 nM) and HG in the presence of the selective PKC beta inhibitors (LY317615, 0.3 μM) for 48 h. Real-time PCR analyses showed mRNA expression of PKC beta and IL-1 beta (a, b). Representative western blots (c) and quantitative determinations of PKC beta and IL-1 beta (d, e) are presented. The data are expressed as the mean ± SD. Experiments were repeated at least three times. NG normal glucose (5.5 mM), HG high glucose (30 mM), PMA (30 nM): phorbol 12-myristate13-acetate; LY (0.3 uM): LY317615; *P < 0.05 vs.NG, **P < 0.01 vs. NG, #P < 0.05 vs. HG or PMA Image collected and cropped by CiteAb from the following publication (//www.cardiab.com/content/15/1/42), licensed under a CC-BY license. Not internally tested by R&D Systems.

The influence of blocking IL-1 beta treatment on the protein expressions of CD31, FSP1, a-SMA, and IL-1 beta . (a–f) HAECs were incubated for 48 h with anti-IL-1 beta antibodies (1000 ng/ml) in the presence of NG or HG. (a1–f1) We performed gene-silencing experiments using transfection with siRNA specific for IL-1 beta . The protein expressions of IL-1 beta , CD31, FSP1 and alpha -SMA were assessed by western blotting. The data are expressed as the mean ± SD. Experiments were repeated at least three times. NG normal glucose (5.5 mM), HG high glucose (30 mM). Anti-IL-1 beta : anti-IL-1 beta antibodies (1000 ng/ml). *P < 0.05 vs. NG or anti-IL-1 beta , #P < 0.05 vs. HG or HG +Vehicle Image collected and cropped by CiteAb from the following publication (//www.cardiab.com/content/15/1/42), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	MAB601-SP
Carrier Free	MAB601-500

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AF-501-NA

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

(67 Publications)

100 μg

AF-501-NA

Recombinant Rat IL‑1 beta /IL‑1F2 (Catalog # 501-RL) stimulates proliferation in the the D10.G4.1 mouse helper T cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Rat IL‑1 beta /IL‑1F2 (10 ng/mL) is neutralized (green line) by increasing concentrations of Rat IL‑1 beta /IL‑1F2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-501-NA). The ND50 is typically 2-10 µg/mL.

IL‑1 beta /IL‑1F2 was detected in immersion fixed LPS-stimulated rat splenocytes using 5 µg/mL Rat IL‑1 beta /IL‑1F2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF‑501‑NA) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained (green). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Spatiotemporal analysis of retinal Il-1 beta protein levels following LD.A-G: Immunohistochemical assessment of Il-1 beta expression (green) in retinal cryosections over the course of LD. A: Immunoreactivity (IR) for Il-1 beta protein was not observed in dim-reared animals. B-D: Il-1 beta -IR was present among ramified nuclei situated within the ONL and OS (arrowheads) immediately following exposure to 24hrs LD. E-F: Il-1 beta -IR co-localised with IBA1+ cells (red) situated in the ONL/OS, though was not apparent in IBA1+ cells outside the vicinity of the ONL (asterisks). G-H: Il-1 beta -expressing cells (H, arrowheads) did not show any discernible IR for the M2 marker CD206 (red). I: Negative control sections, in which the primary antibody was omitted, did not show any resemblance to the IR for Il-1 beta evidenced in C-D at 24hrs LD. J: ELISA for Il-1 beta protein indicated an increased abundance of the protein immediately after 24hrs LD (P0.05). C, choroid; GCL, ganglion cell layer; INL, inner nuclear layer; IPL, inner plexiform layer; IHC, immunohistochemistry; ONL, outer nuclear layer; OS, outer segments; RPE, retinal pigment epithelium. The trend in ELISA protein levels was significant by ANOVA (P 0.05). For CD206+/Il-1 beta - cells, there was no change in their proportion at 24hrs LD (P>0.05). At 3 days post-exposure however the proportion of CD206+/Il-1 beta - cells had tripled (P0.05). C, choroid; GCL, ganglion cell layer; INL, inner nuclear layer; IPL, inner plexiform layer; IHC, immunohistochemistry; ONL, outer nuclear layer; OS, outer segments; RPE, retinal pigment epithelium. The trend in ELISA protein levels was significant by ANOVA (P 0.05). C, choroid; GCL, ganglion cell layer; INL, inner nuclear layer; IPL, inner plexiform layer; IHC, immunohistochemistry; ONL, outer nuclear layer; OS, outer segments; RPE, retinal pigment epithelium. The trend in ELISA protein levels was significant by ANOVA (P 0.05). C, choroid; GCL, ganglion cell layer; INL, inner nuclear layer; IPL, inner plexiform layer; IHC, immunohistochemistry; ONL, outer nuclear layer; OS, outer segments; RPE, retinal pigment epithelium. The trend in ELISA protein levels was significant by ANOVA (P < 0.05); N = 3 for each timepoint. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/26630454), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Rt

Species Glossary

Reactivity: Rt

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	BAF501

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF-501-SP

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MAB201

IL-1 beta/IL-1F2 Antibody (8516) [Unconjugated]

(63 Publications)

100 μg

MAB201

IL‑1 beta /IL‑1F2 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Human IL‑1 beta /IL‑1F2 Monoclonal Antibody (Catalog # MAB201) 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.

IL-1 beta /IL-1F2 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) treated with LPS and monensin using Human IL-1 beta /IL-1F2 Monoclonal Antibody (Catalog # MAB201) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (yellow; Catalog # NL007) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Hsp27 modulates IL-1 beta production in monocytes.(a) Immunoblot analysis of Hsp27 protein expression in classical (CL) and non-classical (NC) monocytes (left), and relative band densities from 2 combined experiments (right), with GAPDH as loading control. (b) Representative immunoblot of Hsp27 and pro-IL-1 beta (p31) in lysates from siRNA-transfected total monocytes for si-control (scrambled control siRNA) or si-Hsp27 (siRNA to Hsp27) either untreated or stimulated with LPS for 3 h. Data in (a) and (b) are cropped images and the corresponding full-length images are presentated in Supplementary Fig. 3. (c) Expression of IL-1 beta mRNA in siRNA-transfected total monocytes (n = 3). (d) Mature IL-1 beta release measured by ELISA of supernatants from siRNA-transfected total monocytes upon LPS stimulation in the presence or absence of BzATP (n = 4). (e) Fold change in IL-1 beta production by LPS and BzATP-treated total monocytes transfected with si-Hsp27, with respect to si-control. Data plotted are mean ± SEM, student t-test (a and b) and two-way ANOVA with Tukey’s multiple comparison test (c), *p < 0.05, **p < 0.01. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27976724), licensed under a CC-BY license. Not internally tested by R&D Systems.

Non-classical monocytes secrete less mature IL-1 beta compared to other monocyte subsets following LPS and BzATP stimulation.(a) IL-1 beta ELISA on supernatants from monocyte subsets either untreated or stimulated with LPS for 24 h in the presence or absence of the P2X7 antagonist, A438079, followed by a secondary stimulation by BzATP in some conditions as indicated. Data plotted are mean ± SEM; n = 8. ****p < 0.0001 ***p < 0.001. ##p < 0.01 and #p < 0.05 is comparison to the respective classical and intermediate subsets treated with LPS + BzATP. Statistics are calculated based on Two-way ANOVA with Tukey’s multiple comparison test (*p < 0.05). (b) Representative cropped immunoblot images for mature 17 kDa (p17) IL-1 beta in culture supernatants of monocyte subsets stimulated as described in (a). The samples were derived from the same experiment and were ran on the same gel. Full-length images are shown in Supplementary Fig. 3. (c) YoPro-1 dye uptake was measured by flow cytometry to reflect level of P2X7R activity, and is shown as the fold change of BzATP-induced YoPro-1 uptake normalized to the respective untreated subset uptake. Data plotted are mean ± SEM; n = 6 based on one-way ANOVA with Tukey’s multiple comparison test. CL: classical, INT: intermediate, NC: non-classical. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27976724), licensed under a CC-BY license. Not internally tested by R&D Systems.

Immunohistochemistry for CD3, IL-1 beta and CD79 expression.Some expression for these markers were found in the CS and CSVT groups at 21 days (positive signal by brown staining). The same overall expression to the materials was seen at the other time points. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0136514), licensed under a CC-BY license. Not internally tested by R&D Systems.

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	FAB201U
Alexa Fluor 350	IC201U-100UG
Alexa Fluor 405	FAB201V
Alexa Fluor 405	IC201V-100UG
Alexa Fluor 488	IC201G-100UG
Alexa Fluor 488	FAB201G
Alexa Fluor 532	FAB201X
Alexa Fluor 594	FAB201T
Alexa Fluor 594	IC201T-100UG
Alexa Fluor 647	IC201R-100UG
Alexa Fluor 647	FAB201R
Alexa Fluor 700	IC201N-100UG
Alexa Fluor 700	FAB201N
Alexa Fluor 750	IC201S-100UG
Alexa Fluor 750	FAB201S
Allophycocyanin	FAB201A
Allophycocyanin	IC201A
Biotin	FAB201B
DyLight 350	FAB201D
DyLight 405	FAB201E
DyLight 488	FAB201K
DyLight 550	FAB201L
DyLight 594	FAB201M
DyLight 650	FAB201W
DyLight 680	FAB201Y
DyLight 755	FAB201Z
Fluorescein	IC201F
HRP	FAB201H
Janelia Fluor 549	FAB201I
Janelia Fluor 646	FAB201J
PerCP (Peridinin-chlorophyll Protein Complex)	IC201C
Phycoerythrin	IC201P
mFluor Violet 450 SE	FAB201MFV450
mFluor Violet 500 SE	FAB201MFV500
mFluor Violet 610 SE	FAB201MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	IC201C
Carrier Free	IC201F
Carrier Free	IC201P
Carrier Free	IC201A
Carrier Free	MAB201-SP
Carrier Free	MAB201-500
Carrier Free	IC201V-100UG
Carrier Free	IC201U-100UG
Carrier Free	IC201T-100UG
Carrier Free	IC201S-100UG
Carrier Free	IC201R-100UG
Carrier Free	IC201N-100UG
Carrier Free	IC201G-100UG

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MAB401

IL-1 beta/IL-1F2 Antibody (30311) [Unconjugated]

(54 Publications)

100 μg

MAB401

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

,

Applications: ,

Host:

Rat Monoclonal

Host: Rat Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	MAB401-SP
Carrier Free	MAB401-500

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AB-401-NA

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

(25 Publications)

1 mg

AB-401-NA

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

, ,

Applications: , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Conjugate:

Unconjugated

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NBP1-19775

IL-1 beta/IL-1F2 Antibody - BSA Free

(21 Publications)

0.1 mg

NBP1-19775

Immunohistochemistry: IL-1 beta/IL-1F2 Antibody [NBP1-19775] - Staining in the cytoplasm of villi and in the crypt cells of human lung carcinoma tissue.

In situ expression of IL-6, IL-1beta, and IL-4 in glomerular and tubulointerstitial compartments in patients with diabetic nephropathy (DN) and control group. D) IL-1beta immunostaining in control and DN groups. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32723296), licensed under a CC-BY licence.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

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 350	NBP1-19775AF350
Alexa Fluor 405	NBP1-19775AF405
Alexa Fluor 488	NBP1-19775AF488
Alexa Fluor 532	NBP1-19775AF532
Alexa Fluor 594	NBP1-19775AF594
Alexa Fluor 647	NBP1-19775AF647
Alexa Fluor 700	NBP1-19775AF700
Alexa Fluor 750	NBP1-19775AF750
Biotin	NBP1-19775B
DyLight 350	NBP1-19775UV
DyLight 405	NBP1-19775V
DyLight 488	NBP1-19775G
DyLight 550	NBP1-19775R
DyLight 594	NBP1-19775DL594
DyLight 650	NBP1-19775C
DyLight 680	NBP1-19775FR
DyLight 755	NBP1-19775IR
FITC	NBP1-19775F
HRP	NBP1-19775H
Janelia Fluor 549	NBP1-19775JF549
Janelia Fluor 646	NBP1-19775JF646
mFluor Violet 450 SE	NBP1-19775MFV450
mFluor Violet 500 SE	NBP1-19775MFV500
mFluor Violet 610 SE	NBP1-19775MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP1-19775SS

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NBP1-42767

IL-1 beta/IL-1F2 Antibody

(1 Review)

(10 Publications)

0.1 mg

NBP1-42767

Immunohistochemistry: IL-1 beta/IL-1F2 Antibody [NBP1-42767] - Analysis of Mouse Embryonic Kidney Tissue: Mouse Embryonic Kidney Fixation: FFPE buffered formalin 10% conc Ag Retrieval: Heat, Citrate pH 6.2. Pressure Cooker Primary antibody: 2ug/ml 1.5 hour @ room T Secondary Ab: MACH 1 HRP POLYMER 1:50 45C RT

Western Blot: IL-1 beta/IL-1F2 Antibody [NBP1-42767] - Analysis using the Biotin conjugate of NBP1-42767. Detection of Lane 1: Mouse IL-1beta Recombinant Protein. Load: 50 ng per lane. Used at a dilution of 1:1,000 overnight at 4C.

Western Blot: IL-1 beta/IL-1F2 Antibody [NBP1-42767] - Detects 10% of the non-denatured (native) precursor 31,000 MW mouse IL-1Beta containing samples but will primarily detect all of the 17,000 MW mature molecule. However, in western blot analysis, the usual procedure of heating the sample in SDS with or without reducing agents will facilitate denaturing of the 31,000 MW IL- 1Beta precursor molecule. Denatured IL-1Beta will have a 18 kDa band.

Immunohistochemistry of Rabbit anti-IL-1 beta/IL-1F2 Antibody in Mouse Embryonic Kidney Tissue: Mouse Embryonic Kidney Fixation: FFPE buffered formalin 10% conc Ag Retrieval: Heat, Citrate pH 6.2. Pressure Cooker Primary antibody: 2ug/ml 1.5 hour @ room T Secondary Ab: MACH 1 HRP POLYMER 1:50 45

This antibody will recognize 10% of the non-denatured (native) precursor 31,000 MW mouse IL-1beta containing samples but will primarily detect all of the 17,000 MW mature molecule. However, in western blot analysis, the usual procedure of heating the sample in SDS with or without reducing agents will facilitate denaturing of the 31,000 MW IL- 1beta precursor molecule. Denatured IL-1beta will have a 18 kDa band.

Reactivity:

Mu, Rt

Species Glossary

Reactivity: Mu, Rt

Applications:

, , , , , , ,

Applications: , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP1-42767AF350
Alexa Fluor 405	NBP1-42767AF405
Alexa Fluor 488	NBP1-42767AF488
Alexa Fluor 532	NBP1-42767AF532
Alexa Fluor 594	NBP1-42767AF594
Alexa Fluor 647	NBP1-42767AF647
Alexa Fluor 700	NBP1-42767AF700
Alexa Fluor 750	NBP1-42767AF750
Allophycocyanin	NBP1-42767APC
Biotin	NBP1-42768
Biotin	NBP1-42767B
DyLight 350	NBP1-42767UV
DyLight 405	NBP1-42767V
DyLight 488	NBP1-42767G
DyLight 550	NBP1-42767R
DyLight 594	NBP1-42767DL594
DyLight 650	NBP1-42767C
DyLight 680	NBP1-42767FR
DyLight 755	NBP1-42767IR
FITC	NBP1-42767F
HRP	NBP1-42767H
HRP	NBP1-42769
Janelia Fluor 549	NBP1-42767JF549
Janelia Fluor 646	NBP1-42767JF646
PE	NBP1-42767PE
PerCP	NBP1-42767PCP
mFluor Violet 450 SE	NBP1-42767MFV450
mFluor Violet 500 SE	NBP1-42767MFV500
mFluor Violet 610 SE	NBP1-42767MFV610

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NBP2-27345

IL-1 beta/IL-1F2 Antibody (43N3D8) - BSA Free

(1 Publication)

0.1 mg

NBP2-27345

Western Blot: IL-1 beta/IL-1F2 Antibody (43N3D8) [NBP2-27345] - Analysis of full length human IL-1 beta recombinant protein (10ng/well) using IL1 beta Antibody (NBP2-27345) at 1:1000 (left lane) and 1:100 (right lane).

Immunohistochemistry-Paraffin: IL-1 beta/IL-1F2 Antibody (43N3D8) [NBP2-27345] - Analysis of Interleukin-1 beta in human kidney tissue using Interleukin-1 beta antibody at 5 ug/ml.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

, ,

Applications: , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Alt Catalog#

IMG-6830A

Alt Catalog# IMG-6830A

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP2-27345AF350
Alexa Fluor 405	NBP2-27345AF405
Alexa Fluor 488	NBP2-27345AF488
Alexa Fluor 532	NBP2-27345AF532
Alexa Fluor 594	NBP2-27345AF594
Alexa Fluor 647	NBP2-27345AF647
Alexa Fluor 700	NBP2-27345AF700
Alexa Fluor 750	NBP2-27345AF750
Allophycocyanin	NBP2-27345APC
Biotin	NBP2-27345B
DyLight 350	NBP2-27345UV
DyLight 405	NBP2-27345V
DyLight 488	NBP2-27345G
DyLight 550	NBP2-27345R
DyLight 594	NBP2-27345DL594
DyLight 650	NBP2-27345C
DyLight 680	NBP2-27345FR
DyLight 755	NBP2-27345IR
FITC	NBP2-27345F
HRP	NBP2-27345H
Janelia Fluor 549	NBP2-27345JF549
Janelia Fluor 646	NBP2-27345JF646
PE	NBP2-27345PE
PerCP	NBP2-27345PCP
mFluor Violet 450 SE	NBP2-27345MFV450
mFluor Violet 500 SE	NBP2-27345MFV500
mFluor Violet 610 SE	NBP2-27345MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP2-27345SS
Azide and BSA Free	NBP2-80793

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MAB4012

IL-1 beta/IL-1F2 Antibody (B122) [Unconjugated]

(12 Publications)

100 μg

MAB4012

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

, ,

Applications: , ,

Host:

Hamster Monoclonal

Host: Hamster Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	MAB4012-SP

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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

Modification

Clonality

Clone

Gene Symbol

Images Type

Isotype

Preservative

Research Areas

Has Sample Size

Promotional Item

Species Glossary

IL-1 beta/IL-1F2 Antibodies

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

IL-1 beta/IL-1F2 Antibody

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

IL-1 beta/IL-1F2 Antibody (2805) [Unconjugated]

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

IL-1 beta/IL-1F2 Antibody (8516) [Unconjugated]

IL-1 beta/IL-1F2 Antibody (30311) [Unconjugated]

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

IL-1 beta/IL-1F2 Antibody - BSA Free

IL-1 beta/IL-1F2 Antibody

IL-1 beta/IL-1F2 Antibody (43N3D8) - BSA Free

IL-1 beta/IL-1F2 Antibody (B122) [Unconjugated]