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LC3B Antibodies

We offer LC3 antibodies that have been cited in 1,150+ publications with 60+ customer reviews. LC3B antibodies include polyclonal and recombinant rabbit monoclonal antibodies. Anti-LC3 antibody validated applications include Knockout-validated Western blot, ELISA, Flow Cytometry, ICC, IHC, IP, and PLA. Our LC3B antibodies have been tested in a variety of model species including Human, Mouse, Rat, Bovine, Canine, Hamster, Porcine, Primate, and Zebrafish. Each LC3 Antibody is fully covered by our 100% Guarantee+. Choose the LC3 antibody best suited for your research needs by viewing technical details, images, citations and reviews.

Alternate Names for LC3B Antibodies

anti-LC3B antibody, anti-MAP1LC3B antibody, anti-ATG8F antibody, anti-Autophagy-related protein LC3 B antibody, anti-Autophagy-related ubiquitin-like modifier LC3 B antibody, anti-lc3-i, ii antibody, anti-lc3b autophagy marker antibody, anti-MAP1 light chain 3-like protein 2 antibody, anti-MAP1A/1BLC3 antibody, anti-MAP1A/MAP1B LC3 B antibody, anti-microtubule associated protein 3 b antibody, anti-microtubule associated protein 3 beta antibody, anti-microtubule-associated protein 1 light chain 3 betaMAP1A/MAP1B light chain 3 B antibody, anti-microtubule-associated proteins 1A/1B light chain 3 antibody, anti-microtubule-associated proteins 1A/1B light chain 3B antibody

7 Results for "lc3b" in Primary Antibodies

NB100-2220

LC3B Antibody - BSA Free

(51 Reviews)

(1669 Publications)

0.1 ml

NB100-2220

Lysates of HeLa parental cell line and LC3B knockout HeLa cell line (KO) untreated (-) or treated (+) with 50 uM CQ for 18 hours. PVDF (Polyvinylidene difluoride) membrane was probed with 0.5 ug/mL of Rabbit Anti-LC3B Polyclonal Antibody (Catalog # NB100-2220) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog# HAF008). A specific band was detected for LC3B at a molecular weight of approximately 15 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. GAPDH is shown as a loading control. This experiment was conducted under reducing conditions.

<b>Genetic Strategies Validation. </b>LC3B was detected in immersion fixed CQ treated HeLa cells (left) but was not detected in LC3B knockout HeLa cells (right) using rabbit anti-human LC3B polyclonal antibody (Catalog #NB100-2220) at 0.3 ug/mL for 3 hours at room temperature. Cells were stained using the NorthernLights(TM) 557-conjugated anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm.

<b>Biological Strategies Validation. </b>Contribution of autophagy to lipid droplet formation in WT and KO NSC34 cells.(A) WT and KO NSC 34 cells were treated with (+) or without (-) bafilomycin A (bafA) under basal or HBSS starvation conditions for 4 h and blotted for p62, LC3, and GAPDH. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32321733) licensed under a CC-BY license.

TMEM16A inhibited AngII-induced autophagy in aortas. (D) Representative immunofluorescence staining of LC3 (green) and a-SMA (red) in aortas. Scale bars, 20 um. Quantification of LC3B puncta. Five random fields (2,500 um2/filed) were measured in one section. **P < 0.01 vs. sham + TMcon; ##P < 0.01 vs. AngII + TMcon, one-way ANOVA. n = 14 sections from seven mice per group. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32226533) licensed under a CC-BY license.

LC3 accumulation in muscle fibers from patients with PAD (row B - E). Normal fibers from a Non-PAD sample (row A). Scale bar = 100 uM. Image from verified customer review.

Localization of LC3 by immunohistochemical method. The immunohistochemical staining shows that LC3, CRF, and HIF-1alpha have an overlapping localization in villous and extravillous trophoblast. d: decidua; v: villi; DeVe: decidual vessel. Original magnification 40x. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/bmri/2013/689768/) licensed under a CC-BY license.

Detection of LC3B in mouse ES cell lysates using Rabbit anti- [Catalog # NB100-2220]. LC3 detected at a molecular weight of approximately 17 kDa. Atg5-/- ES cells from Dr. Noboru Mizushima [Mizushima, N. et al. J. Cell Biol. 152 (2001)] Photo courtesy of Dr. Beth Levine, UT SW Medical Center.

ORP1L and VAP-A form cholesterol-dependent ER-AV contact sites MelJuSo cells expressing GFP or GFP-tagged ORP1L mutants were fixed and stained for LC3 and Calnexin. Y477 and D478 in the FFAT motif were mutated to alanines in the ORP1L ydaa mutant. Scale bar, 10 um. Right: co-immunoprecipitation for ORP1L (mutants) with VAP-A. GFP-ORP1L mutants or GFP were isolated from lysates of HEK293T cells co-overexpressing HA-VAPA using GFP-Trap beads. Western blot filters were probed for isolated GFP-tagged proteins, the associated HA-VAP-A and the input HA-VAP-A, as indicated. Image collected and cropped by CiteAb from the following publication (//www.nature.com/doifinder/10.1038/ncomms11808), licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>Lysates of mouse NIH3T3 and rat PC-12 cell lines untreated (-) or treated (+) with CQ. PVDF (Polyvinylidene difluoride) membrane was probed with 0.5 ug/mL rabbit anti-LC3B polyclonal Antibody (Catalog # NB100-2220, Novus Biologicals), followed by 1:2000 dilution of goat anti-rabbit IgG secondary antibody. LC3 detected at a molecular weight of approximately 15 kDa in treated NIH3T3 and PC-12 cells.

<b>Biological Strategies Validation. </b>Detection of LC3I and LC3II in mouse cochlea cell line SV-K1 using Rabbit anti- [Catalog # NB100-2220]. Cells were treated with CQ (1 uM), and As2O3 (1 uM) for 24 hrs. Image from verified customer review.

Autophagy Investigation. (A) Representative WBs (Western Blots) of LC3B-I, LC3B-II and GAPDH loading control in DANs derived from 4 iPD patients and 4 healthy controls. (B) WBs from three independent experiments were quantified by densitometry and LC3B-II normalized to a loading control. Grouping of data by healthy vs iPD as well as iPD stratified for T/T and C/C genotype. (C) Ratio of LC3B-II (normalized to loading control) following bafilomycin treatment/untreated. Citation: Marrone L, Bus C, Schondorf D, Fitzgerald JC, Kubler M, Schmid B, et al. (2018) Generation of iPSCs carrying a common LRRK2 risk allele for in vitro modeling of idiopathic Parkinson's disease. PLoS ONE 13(3): e0192497. //doi.org/10.1371/journal.pone.0192497

<b>Biological Strategies Validation. </b>Expression of autophagic protein LC3 in BME-UV1 cells forming acinar structures on Matrigel. Representative images of Western blot analysis of LC3 in bovine MECs grown in 3D culture for 3, 6, 9, and 14 days in differentiation medium (control), enriched with 17beta-estradiol (E2, 1 nM), progesterone (P4, 5 ng/mL), or both (E + P); expression of beta-actin was used as a loading control; graphs below the images show the results of densitometric analysis, in which IOD of each band was measured, and the values were normalized to IOD of beta-actin; the IOD results are presented as means +/- SEM from at least three separate experiments. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/bmri/2014/382653/) licensed under a CC-BY license.

GNA triggers the formation of autophagic markers in A549 and HeLa cell. Effects of GNA on LC3 protein. A549 cells were treated with various concentrations of GNA for 24 hours or 3 uM GNA for the indicated periods of time, then analyzed by western blotting using anti-LC3 antibodies. GAPDH protein was used as the loading control. The bar graph shows the band intensities of LC3-II relative to those of GAPDH. Mean+/- SEM, n = 3, *means p<0.05, ** p<0.01, one-way ANOVA. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0083604), licensed under a CC-BY license.

Autophagosomes accumulate in neurons of the VH gray matter at day 1 after SCI. Representative images of IHC staining for LC3 (green) and neuronal marker NeuN (red) in VH of gray matter from sham and SCI animals. Stronger co-localization between NeuN and LC3 is apparent at day 1 after SCI. Scale bar is 20 um. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2014527) licensed under a CC-BY license.

Autophagosomes accumulate in microglia and oligodendrocytes of the dorsal white matter adjacent to injury after SCI. Representative images of IHC staining for LC3 (green) and activated microglia marker CD11B (red) in dorsal white matter of sham and SCI animals. Increased co-localization between LC3 and CD11B is apparent at day 1 after SCI. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2014527) licensed under a CC-BY license.

Staining of rat brain tissue section. Analysis using Rabbit anti-LC3B Antibody [Catalog # NB100-2220]. Image from verified customer review.

Human ovarian Cancer tissue stained using heat mediated antigen retrieval in pH 6.0 citrate buffer at 1:200 dilution. Image provided by verified customer review.

FFPE (Formalin-Fixed Paraffin-Embedded) tissue section of mouse brain using 1:200 dilution of Rabbit anti-LC3B antibody [Catalog # NB100-2220]. The specific signal of LC3 was detected using HRP-conjugated secondary antibody with DAB (3, 3 -diaminobenzidine) reagent, and nuclei of cells were counterstained using hematoxylin. This LC3B antibody generated a low to moderate levels of cytoplasmic staining in the glial cells. The neurons depicted a moderate to strong staining for LC3 in their cytoplasm.

Analysis using the Biotin conjugate of Rabbit anti-LC3B Antibody [Catalog # NB100-2220]. Staining of brain, cerebral cortex, neurons with cell processes.

SCI leads to lysosomal dysfunction. Representative high magnification images ( x 60) of LC3 (green) and CTSD (red) staining in VH of gray matter from sham and day 1 SCI animals. Decrease in number and intensity of CTSD puncta (lysosomes) and increase in LC3 puncta (autophagosomes) is apparent after SCI. Scale bar is 10 um. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2014527) licensed under a CC-BY license.

Image shows a specific band for LC3B at a molecular weight of approximately 15 kDa in 0.5 mg/mL of Neuro2A lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

LILRB2 antagonism effects on the bactericidal function of the MDSCs are independent of autophagy. (B) Lysates of cells from panel A were collected at 18 h and analyzed using Western blot for the autophagy marker, microtublue-associated light chain-3 (LC3). Lipidation is indicated by LC3-II. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35757769), licensed under a CC-BY licence.

$CQ-induced autophagy inhibition leads to Nrf2 pathway activation in C2C12 muscle cells. a. Treatment of differentiated C2C12 myotubes with autophagy inhibitor CQ increased the sarcoplasmic level of LC3-II and SQSTM1 proteins in a dose- and time-dependent manner, indicating effective inhibition of the autophagic flux. Autophagy inhibition was accompanied by a slight decrease in the sarcoplasmic level of the soluble Keap1 protein (possibly reflecting its sequestration into insoluble protein aggregates) and an increase in the nuclear level of Nrf2 protein. GAPDH and lamin B were used as loading controls for sarcoplasmic and nuclear protein-enriched fractions, respectively; a representative of 3 independent experiments is shown. Image collected and cropped by CiteAb from the following publication (//actaneurocomms.biomedcentral.com/articles/10.1186/s40478-016-0384-6), licensed under a CC-BY licence.$

Accumulation of an autophagosome marker in the presynaptic terminals of NMJs of SOD1G93A mice (A) Representative confocal micrographs of immunohistochemistry showing accumulation of the autophagosome marker LC3 (grayscale in left panels, green in right panels) in presynaptic terminals of diaphragm NMJs in SOD1G93A mice at P120, but not at NMJs of wild-type mice at P140. Nerves were stained using anti-neurofilament and anti-SV2 antibodies (blue), and postsynaptic acetylcholine receptors were labeled with Alexa 594-conjugated alpha-bungarotoxin (red). In these cross-section images of NMJs, presynaptic terminals are above the bungarotoxin signal, and postsynaptic myotubes are below the bungarotoxin signal. Scale bar: 10 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28890682), licensed under a CC-BY licence.

CAIX expression in PDAC and association with outcomeA. Representative images of immunohistochemical staining showing: negative expression of CAIX in both tumor cell and stromal compartments (top left), positive expression of CAIX in tumor cells but not stromal compartment (top right), positive expression of CAIX in stromal compartment but not tumor cells (bottom left), positive expression of CAIX in both tumor cell and stromal compartments (bottom right). B. Kaplan-Meier analysis of CAIX expression in the distinct tumor cell and stromal compartments and the associated overall survival. C. Representative images of immunohistochemical staining showing low stromal volume and high stromal volume and low and high microvessel density D. Representative images of immunohistochemical staining showing: positive expression of MCT4 in tumor cell compartment and positive expression of MCT4 in tumor stromal compartment. Kaplan-Meier analysis of combinatorial MCT4 and CAIX expression in the stromal compartment and the associated overall survival. Hazard ratios and p-values for each comparison are summarized in the table. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27623078), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Al, Av, Ba, Bv, Ca, Ch, ChHa, Gp, Ha, In, Pm, Pm, Rb, SyHa, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Al, Av, Ba, Bv, Ca, Ch, ChHa, Gp, Ha, In, Pm, Pm, Rb, SyHa, Ze

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	NB100-2220AF350
Alexa Fluor 405	NB100-2220AF405
Alexa Fluor 488	NB100-2220AF488
Alexa Fluor 532	NB100-2220AF532
Alexa Fluor 594	NB100-2220AF594
Alexa Fluor 647	NB100-2220AF647
Alexa Fluor 700	NB100-2220AF700
Alexa Fluor 750	NB100-2220AF750
Allophycocyanin	NB100-2220APC
Biotin	NB100-2220B
DyLight 350	NB100-2220UV
DyLight 405	NB100-2220V
DyLight 488	NB100-2220G
DyLight 550	NB100-2220R
DyLight 594	NB100-2220DL594
DyLight 650	NB100-2220C
DyLight 680	NB100-2220FR
DyLight 755	NB100-2220IR
FITC	NB100-2220F
HRP	NB100-2220H
Janelia Fluor 549	NB100-2220JF549
Janelia Fluor 646	NB100-2220JF646
PE	NB100-2220PE
PerCP	NB100-2220PCP
mFluor Violet 450 SE	NB100-2220MFV450
mFluor Violet 500 SE	NB100-2220MFV500
mFluor Violet 610 SE	NB100-2220MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-2220SS

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NB600-1384

LC3B Antibody - BSA Free

(19 Reviews)

(397 Publications)

0.1 ml

NB600-1384

FTC-133/TPC-1 cells were transfected with HMGB1 shRNA and control shRNA and then pre-treated for 1 h with pepstatin A (PA, 10 uM) and E64D (10 uM) as indicated. Cells were subsequently treated for 3 h with HBSS in continuous presence or absence PA/E64D inhibitors. LC3-I/II levels were assayed by Western blot. Image collected and cropped by CiteAb from the following publication (//jeccr.biomedcentral.com/articles/10.1186/s13046-019-1328-3) licensed under a CC-BY license.

Analysis of LC3B in HeLa cells using anti-LC3B antibody (red) [Catalog # NB600-1384]. Nuclei were counterstained with DAPI (blue).

LC3B immunohistochemical staining in CC tissue(A) CC with low LC3B dot like staining. (B) CC with high LC3B dot like staining. (magnification 20X). Image collected and cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.17554) licensed under a CC-BY license.

Analysis of LC3B in treated U87-MG (human glioblastoma astrocytoma) lysates using anti- [Catalog # NB600-1384].

Depletion of RARalpha prevents ATRA-induced autophagic flux in SKBR3 cells. LC3B western blotting and quantification of the autophagic activity in control and RARalpha-knockdown SKBR3 cells upon treatment with 1 uM ATRA for 2 days in the presence or absence of BafA for 2 h at 200 nM. LC3B-II expression was normalized to GAPDH and to vehicle-treated control cells. Standard deviations from at least five independent experiments are shown. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2015236), licensed under a CC-BY license.

Human cervical carcinoma (HeLa) and Mouse Neuroblast cells (Neuro2a) were treated with (+) and without (-) 50 uM Chloroquine overnight. Whole cell protein lysates were prepared in 1x Laemmli sample buffer and approximately 10 ug of each lysate (NBP2-49689 and NBP2-49688) was separated on a 4- 20% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% nonfat milk in TBST. The membrane was probed with 2 ug/mL anti-LC3 (NB600-1384) and 1 ug/mL anti-alpha tubulin (NB100-690) as a loading control, and detected with the appropriate secondary antibodies using NovaLume Pico Chemiluminescence Substrate (NBP2-61915).

<b>Biological Strategies Validation. </b>Fibroblasts were exposed to 1 mM LLOMe and when indicated pretreated with the autophagy inhibitors 3-methyladenine (5 mM, 1 h) or bafilomycin A1 (25 nM, 30 min). Immunostaining with corresponding colocalization analysis of a) LC3 (green) and galectin-3 (red) and b) LC3 (green) and LAMP2 (red). Merged images show colocalization in yellow, nuclei are stained with DAPI (blue). LLOME = l-leucyl-l-leucine methyl ester, a lysosomal membrane damage compound. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32409651/) licensed under a CC-BY license.

Confocal analysis of HeLa cells using Rabbit anti-LC3B antibody (Catalog # NB600-1384, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).

Immunocytochemical/Immunofluorescent staining of treated U373-MG cells using the HRP conjugate of anti- (Catalog # NB600-1384). The nuclei were stained with DAPI.

Staining of treated U373-MG (human glioblastoma) cells using anti- [Catalog # NB600-1384].

Analysis of U87MG glioma xenografts using anti- [Catalog # NB600-1384]. Image from verified customer review.

LC3B staining in treated U87-MG cultured & subcutaneous tumors.

LC3B staining in glioblastoma multiform tissue.

Lane view shows a specific band for LC3B in 0.5 mg/ml of Neuro2A lysate at a molecular weight of approximately 15 kDa. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

LC3A and LC3B siRNAs specifically block the expression of the LC3A and LC3B proteins, respectively, in A549 cell line (E1,2,3). In E4 the reactivity of and of the LC3B (5F10 antibody) is shown, following silencing of the LC3A or of the LC3B genes, in the A549 cell line. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0137675), licensed under a CC-BY license.

NOD1-dependent induction of autophagy contributes to IRGM and LC3 recruitment to Mtb-containing vesicles. The subcellular localization of autophagy proteins was observed in untreated (A, C) and Tri-DAP treated cells (B, D) using anti-IRGM and anti-LC3 antibodies, which were detected using a secondary antibody coupled to 5 nm gold particles (indicated with arrowheads; TEM X 62,000). Gold particles colocalizing with bacteria were manually counted in 10 macrophages of each condition (E, F) and the differences between the treatments are indicated: ***p < 0.01 and *p < 0.05 vs. Mtb, ++p < 0.01 vs. Tri-DAP, Wilcoxon Rank test. Box plots indicate median and quartiles. Image collected and cropped by CiteAb from the following publication (//bmcpulmmed.biomedcentral.com/articles/10.1186/1471-2466-14-152), licensed under a CC-BY license.

Examples of immunohistochemical stainings; (A) LC3B low, (B) LC3B high, (C) p62 low (any cellular compartment), (D) p62 cytoplasmic low, (E) p62 cytoplasmic/dot like high, nuclear low, (F) p62 cytoplasmic/dot like low, nuclear high, (G) HMGB1 low, (H) HMGB1 high. Objective magnification, 40×.

Validation of modulations in autophagy-related markers after SIRT6 knockdown in melanoma cells. (B) Western blot analyses of various autophagy markers including LC3 I and II, BECN1, SQSTM1, ATG3, ATG7, ATG10 and GAA using cell lysates of shNS and shSIRT6 in A375 melanoma cells. beta-actin was used as loading control. The figure shows representative western blot images from 2-3 experiments. Image collected and cropped by CiteAb from the following publication (//www.genesandcancer.com/lookup/doi/10.18632/genesandcancer.153), licensed under a CC-BY licence.

Induction of autophagic flux in OE19 upon Lapatinib treatment. (a) LC3B flux was assessed comparing control and BafilomycinA (BafA)-treated (200 nM, 2 h) OE19 upon Lapatinib treatment (120 nM, 24 h). LC3 band intensities were quantified using ImageJ software. Total protein was used as a loading control, and Phospho-Her2 for Lapatinib treatment (n = 3). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30297650), licensed under a CC-BY licence.

In vitro alphaS seeds induce endo-lysosome-free and endo-lysosome-associated alphaS inclusions in H4/V1S-SV2 cells. (d) The treated cells were further subjected to immunocytochemical staining of LAMP1 (Alexa fluor 405) and LC3 (Alexa fluor 568) to demonstrate the association between autophagosomes, endo-lysosomes, and seeded alphaS inclusions. Scale bar: 5 um (top row) and 2 um (bottom row). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-08149-w), licensed under a CC-BY licence.

(E) LC3A and LC3B siRNAs specifically block the expression of the LC3A and LC3B proteins, respectively, in A549 cell line (E1,2,3). In E4 the reactivity of LC3A (ab62720 Ab) and of the LC3B (5F10 antibody) is shown, following silencing of the LC3A or of the LC3B genes, in the A549 cell line. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26378792), licensed under a CC-BY licence.

Combination treatment of valproic acid (VPA) and doxorubicin (DOX) synergistically augmented the autophagy of HepG2 cells. (D) LC3 I and II protein levels were analyzed using Western blotting. Actin was used as the loading control. The intensity of LC3B-II bands was quantified by scanning densitometry program ImageJ and normalized to that of actin (right panel). Three independent experiments were performed and results reported as the mean +/- standard deviation (SD). ** p < 0.01, *** p < 0.001 compared with the control group. Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/1422-0067/18/5/1048), licensed under a CC-BY licence.

Autophagy and the proteasome regulate CagA stability. (A) Wild-type (WT) and autophagy-deficient (Atg5-/-) MEFs were infected with a CagA+ vacA- isogenic mutant strain (MOI 100) for 8 hours using a gentamycin protection assay. The autophagy marker LC3-II was used to confirm that Atg5-/- MEFs are autophagy-deficient. CagA protein levels were measured by Western blotting using beta-actin as loading control. Graph shows fold change of CagA normalized to beta-actin relative to WT MEFs (mean + SEM; n = 4). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30631092), licensed under a CC-BY licence.

PB induces autophagy but blocks autophagosome trafficking and autophagosome-lysosome fusion in Hep3B cells. (B) Hep3B cells were treated for 6, 24 and 48 h with the indicated concentrations of PB. Levels of protein expression were analysed by Western blot using antibodies against LC3B, p62/SQSTM1, ATG12, BECN1 and Actin. Images were cropped from different blots run under the same experimental conditions. The original blots were attached as Supplementary Figure 3A. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep41437), licensed under a CC-BY licence.

Effects of exogenous HMGB1 on ATRA-induced differentiation. E). Expression of PML-RARalpha, P62 and LC3-I/II in NB4 cells treated with ATRA (1 uM) or HMGB1 (10 ug/ml) for 48 h was assayed by western blotting and co-immunoprecipitation in comparison to control (DMSO). Image collected and cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.15432), licensed under a CC-BY licence.

Activation of autophagy by MG132 is associated with AGR2 decline. d) The levels of AGR2 and LC3B were measured by western blotting with MG132 treatment at indicated time. e Analysis of LC3B and AGR2 accumulation in A549 cells by fluorescent staining. Scale bar = 10 um Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30647455), licensed under a CC-BY licence.

Autophagy was induced under conditions of LH and inhibited by CQ and 3-MA in HCC cells.(A-C). SMMC-7721 and HepG2 cells were directly incubated under normal or LH conditions for 8 h or incubated under LH conditions in the presence or absence of CQ or 3-MA for 8 h. (C). The whole-cell lysates were subjected to western blot analysis as described in the 'Materials and methods'section. The experiments were repeated at least three times. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep05382), licensed under a CC-BY licence.

The effect of H19 siRNA on autophagy in OGD/R model. A) Immunofluorescence of LC3II in different conditions. Image collected and cropped by CiteAb from the following publication (//www.aginganddisease.org/EN/10.14336/AD.2016.0530), licensed under a CC-BY licence.

Validation of selected differentially expressed genes. A. RT-PCR analysis of genes elevated in drug resistant cells. The y-axis represents fold up-regulation in the different drug resistant cell lines over the parental OV90 cell line. B. RT-PCR analysis of genes decreased in drug resistant cells. The y-axis represents the fold down-regulation of the different resistant cell lines compared to the parental OV90 cell line. C. Immunoblot analysis of selected gene products identified by microarray and RT-PCR as altered in drug resistant cells. Image collected and cropped by CiteAb from the following open publication (//ovarianresearch.biomedcentral.com/articles/10.1186/1757-2215-4-21), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Osteogenic differentiation in 3D. (A) Histochemical staining of 12-μm-thick gel sections from one representative donor after 5 weeks of culture in undifferentiated or osteogenic conditions. Osteoimage® (HA-specific staining), von Kossa (stains anionic portion of phosphates, carbonates, and other salts) and Alizarin Red (stains Ca2+ deposits) shows differentiation in the Col I—BMSC cultures. Scale bar = 100 μm. (B) Immunostaining for osteocalcin (OCN) and nuclear 4′,6-diamidino-2-phenylindole (DAPI) in 3D cultures after 5 weeks. (C) Immunostaining for osteopontin (OPN) and nuclear DAPI in 3D cultures after 5 weeks. Scale bar = 50 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34948393), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

G-TPP induces PINK1 stabilization and kinase activity in HeLa cells(A) G-TPP treatment leads to PINK1 stabilization and pS65-Ub induction in HeLa cells. HeLa cells stably expressing untagged Parkin were treated with 10 µM G-TPP for the indicated times. Western blots were prepared with cell lysates and probed with antibodies against PINK1 and pS65-Ub. GAPDH served as a loading control. (B) pS65-Ub is induced in G-TPP treated cells and co-localizes with EGFP-Parkin and mitochondria. HeLa cells stably expressing EGFP-Parkin (green) were treated with 10 µM G-TPP for the indicated times and fixed. Cells were stained with antibodies against pS65-Ub (red) and the mitochondrial marker TOM20 (cyan). Scale bars correspond to 10 µM. (C) Quantification of Parkin translocation using High Content Imaging. HeLa EGFP-Parkin cells were treated for 4 or 8 h with or without 10 µM G-TPP. CCCP treatment (10 µM for 2 h) was used as a positive control. Cells were fixed, counterstained with Hoechst dye to visualize nuclei, imaged and analyzed using the ratio of cytoplasmic to nuclear EGFP signal [21]. Data was normalized to positive (2 h 10 µM CCCP treatment) and negative (2 h DMSO) controls. G-TPP significantly induced Parkin re-localization to levels similar to or beyond 2 h CCCP treatment. Shown are the mean values of three independent experiments with triplicate wells each ± SEM (one-way ANOVA with Tukey’s posthoc, ***p < 0.0005). Image collected and cropped by CiteAb from the following open publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.22287), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Association of p53 and APE1 on p53-binding sites in p21 promoter.(A) p21 promoter structure showing p53 and AP4 binding sites. (B) ChIP Real Time PCR analysis showing relative enrichment (2− delta CT) of APE1-immunoprecipitated DNA over that from control IgG in p21 promoter regions containing p53 binding sites 1 & 2 in HCT116WT cells. (C) Re-ChIP analysis (first IP with alpha -APE1 and the second IP with alpha -p53 antibody) showing simultaneous recruitment of APE1 and p53 in control vs. etoposide treated cells; *: p value <0.05 (n = 2) calculated based on APE1/p53 enriched DNA from control vs. etoposide treated cells. (D) Western analysis of FLAG immunoprecipitate (IP) to detect APE1-associated p53 and FLAG (APE1) from empty vector vs. FLAG-tagged WT APE1 or FLAG-tagged N delta 33 APE1 transfected HCT116WT cells (left panel) and from control vs. etoposide-treated WT APE1-FLAG transfected cells (right panel). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/23874636), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Loss of functional PRPF31 induce RNA:DNA hybrid dependent genomic instability but not in mice retinal neurons.(a andb) gamma H2AX and 53BP1 foci analysis in PRPF31 siRNA-transfected RPE-1 cells. (c and d) gamma H2AX and 53BP1 foci analysis in vasculo-stromal fraction derived primary cells fromPrpf31+/A216P mice (Prpf31-ki). (e) gamma H2AX and 53BP1 foci analysis in retina fromPrpf31+/A216P mice on postnatal day 20. All column bars represent the mean. For (a-d) “n”, mentioned on respective column, signify number of cells analyzed from two independent experiments. For (e) n=16 for each column and signify number of retinal sections analyzed; acquired from n=4 eyes. Error bars represent Standard error of Mean (SEM). *P≤0.05; **P<0.01, ***P<0.001 using Mann-Whitney test (a,b), Kruskal-Wallis test followed by Dunn’s post hoc test (c,d); and two tailed unpaired Student’s t-test (e). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30345028), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Subarachnoid hemorrhage (SAH) grades and temporal expression of endogenous osteopontin (OPN) and autophagy‐related proteins in rat brain after SAH. A, Representative brain images of Sham and SAH rats. B, Summary of SAH grading scores of all groups. Sample size is 27, n = 9 per group. Data were analyzed using Kruskal‐Wallis test, χ2 = 18.183. C, Representative Western blot images and quantitative analyses of OPN, Beclin 1, LC3, and ATG 5 from the left hemisphere of rat brains at different time points after SAH. Sample size is 36, n = 6 per group. Data were presented as mean ± SD. F = 28.45 for OPN, F = 12.37 for Beclin 1, F = 18.88 for LC3, F = 22.14 for ATG5. *P < .05, ** P < .01, ***P < .001 vs Sham group. SAH, subarachnoid hemorrhage; Vehicle, phosphate‐buffered saline; rOPN, recombinant OPN. NS, not significant Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31436915), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Staufen1 co-fractionates with Gag and vRNA in gradient density fractionation analyses. (A) HeLa cells were either mock transfected with empty vector, pcDNA3, or with a plasmid that expresses Staufen1-HA. The transfected cells were collected 24 h later, lysed and were either mock-treated or treated with RNAse A. The lysates were then fractionated on 5–50% sucrose gradients and 20 fractions were collected for further analysis by western blotting for viral and host proteins, as indicated. HIV-1 viral genomic RNA (vRNA, 9 kb) was assessed in each fraction by slot blot analysis. TL represents the total lysates. (B) HeLa cells were co-transfected with pNL 4–3 and Staufen1-HA. The presence of Staufen1-HA, precursor Gag and p24 were assessed in each fraction by western blotting analysis. HIV-1 viral genomic RNA (9 kb, vRNA) was assessed in each fraction by slot blot analysis. Staufen1, Gag and vRNA were quantitated in each fraction by densitometry and relative levels are depicted for each fraction (Blue: Staufen1-HA, Red: Gag, Green-vRNA). Image collected and cropped by CiteAb from the following open publication (//journal.frontiersin.org/article/10.3389/fmicb.2012.00367/abstract), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Endogenous MIR376A limits starvation-induced autophagy.(A) Blockage of endogenous MIR376A by Ant-376a, but not CNT-Ant further stimulated starvation (STV)-activated LC3-I to LC3-II conversion in MCF-7 cells. ACTB was used as a loading control. LC3-II/LC3-I densitometric ratios were marked. (B) Ant-376a, but not CNT-Ant resulted in further activation of SQSTM1 protein degradation following starvation in MCF-7 cells. SQSTM1/ACTB ratios were marked. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/24358205), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

GAS6 increases the amount of both CIP2A and p-DAPK in RSC96 cells(A) RSC96 cells were incubated with GAS6 (100 ng/ml), and immunoblotting evaluations of CIP2A and p-DAPK expression were performed. The lower panel shows the intensity during basal conditions from the scanned imaged (upper panel). (B) RSC96 cells incubated with and without GAS6 (100 ng/ml) for 30 min and analysed by immunofluorescence. (C) Immunohistochemistry images demonstrated that CIP2A expression levels were higher than sham levels at 14 days and 28 days in the cavernous nerve after BCNI. Image collected and cropped by CiteAb from the following open publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.23978), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Effect of treatment with FX extract on CREB/BDNF signaling. (a-c) Differences in the phosphorylation of CREB and expression of BDNF between groups in the PFC and hippocampus (n = 3-4). (d, e) pCREB and BDNF immunofluorescence was assessed in the PFC and hippocampus (n = 3-4). Mean ± SD. #P < 0.05 versus normal group; ∗P < 0.05 and ∗∗P < 0.01 versus control group. Scale bar = 200 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30065945), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry of seven in absentia homolog 2 (SIAH2) gene in normal, in situ and invasive breast carcinomas. (A) Occasional nuclear positivity (arrows) in luminal cells in the terminal duct lobular unit. (B) Moderate to strong staining of SIAH2 in the nucleus of a small proportion of the cell in a high nuclear grade ductal carcinoma in situ with comedo necrosis. (C) Occasional weak to moderate SIAH2 (arrows) staining in a luminal type ductal carcinoma. (D) Strong SIAH2 staining in all nuclei in this basal-like breast carcinoma. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/21306611), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

CNF1-triggered IL-1 beta maturation requires activated Rac, ASC and caspase-1.(A) Western blot analysis of the production and maturation/secretion of IL-1 beta by primary macrophages following treatment with CNF1, LPS or CNF1+LPS for 10 h. Actin and BSA were used as loading controls. (B) Quantification of caspase-1 activity in macrophages following treatment with CNF1+LPS for 6 h using YVAD-Fluorescent Labelled Inhibitor Caspase-1 Activity (FLICA). (C) Western blot analysis of macrophages IL-1 beta maturation/secretion upon transfection of HA-Rac2Q61E and LPS treatment. (D) Co-immunoprecipitation of Myc-Rac2 and caspase-1 using an anti-Myc antibody following the treatment of HEK 293T cells with CNF1+LPS for 6 h. Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.ppat.1004732), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Expression of sphingosine-1-phosphate (S1P), interleukin (IL)-1 beta , and protease-activated receptor-1 (PAR-1) in isolated astrocytes. (A) Isolated and purified astrocytes were identified by bright-field microscopy and immunofluorescence for glial fibrillary acid protein (GFAP; red). Nuclei (DAPI) were labeled in blue. Scale bar = 100 μm. (B) Western blot and quantitative reverse transcription polymerase chain reaction (qRT-PCR) of the protein and mRNA expression, respectively, of S1P 0, 1, and 6 h after astrocytes were treated with lipopolysaccharide (LPS) or thrombin and Dabigatran (Dab) or PAR-1-inh (LPS only). (D) qRT-PCR and (E) enzyme-linked immunosorbent assay (ELISA) of the mRNA expression and secretion, respectively, of IL-1 beta 0, 1, and 6 h after astrocytes were treated with LPS or thrombin and Dab or PAR-1-inh (LPS only). (F) Immunofluorescence of astrocytes and quantification of relative mean integrated optical density (IOD; green fluorescence). PAR-1 was stained in green and nuclei (DAPI) were stained in blue. Scale bar = 100 μm. For (B–F), data are presented as the mean ± SD (n = 3). *p < 0.05; #p < 0.05 vs. the same group at 0 h; &p < 0.05 vs. the same group at 1 h. ns, not significant. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32694981), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

53BP1/RIF1 work epistatically to counteract BRCA1/CtIP-mediated end resection to regulate DSB repair pathway choiceA. HeLa cells harboring pGC were treated with the indicated siRNAs before transfection with I-SceI-expressing vector to induce DSBs. After 48h, the percentage of GFP+ cells was measured as an indication for HR efficiency. B.-D. Asynchronous A549 cells were treated with the indicated siRNAs before irradiation with 2Gy and CtIP B. and RPA foci C. were monitored at 2h while RAD51 D. foci were enumerated at 4h. E. Left panel: representative photos for the colocalization between 53BP1 and RIF1 foci in A549 cells at 2h after 2Gy. Right panel: quantitation of 53BP1, RIF1 or colocalized foci in the absence (DMSO) or presence of ATM inhibitor (ATMi). F. Left panel: representative micrographs for 53BP1 or RIF1 foci in irradiated cells after depletion of either proteins. Right panel: quantitation of experiments presented in the left panel. At least 100 nuclei were counted. In all cases, the number of foci measured in non-irradiated cells was subtracted (relative). Shown are the mean ±SEM for three independent experiments. sc: scrambled RNA, si53: si53BP1, siR: siRIF1, siB: siBRCA1, and siC: siCtIP. Image collected and cropped by CiteAb from the following open publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.11023), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Up-regulation of RAS induced by cholesterol and 25-hydroxycholesterol in HK-2 cells. HK-2 cells were made quiescent by serum-free medium for 24 hours and then maintained in serum-free medium (control) or serum-free medium containing 30 μg/ml cholesterol together with 1 μg/ml 25-hydroxycholesterol (lipid) for 24 hours. (A) Real-time PCR of the total RAS components mRNA prepared from HK-2 cells with or without lipid treatment. AT1/AT2 ratio was evaluated as the balance between AT1 and AT2. beta -actin was used as mRNA loading control. (B) Western blot analysis for RAS components protein expression. (C) The histogram shows the average volume density corrected by the housekeeping control, beta -actin. Data is expressed as mean ± SD. *P < 0.05 vs. control. Angiotensinogen, AGT; angiotensin II, Ang II; angiotensin converting enzyme, ACE; angiotensin II type 1 receptor, AT1; angiotensin II type 2 receptor, AT2. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/23570453), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Effect of TERT silencing on murine M1 macrophage polarization.TERT siRNA and GV144-TERT were transiently transfected into LPS-treated RAW264.7 cells, respectively. (a) The endogeous TERT levels were detected by real-time PCR and western blot. The results are shown as relative expression against control expression without treatment. (b) The mRNA levels of M1 macrophages biomarkers including TNF-alpha , IL-1 beta , NOS2 and CCL2 were detected by real-time PCR. The results are shown as relative expression against control expression without treatment. (c) The secretion of proinflammatory cytokines including TNF-alpha , IL-1 beta , IL-6 and IL-12 were determined by ELISA. (d) TERT successful over-expression was verified by real-time PCR and western blot in LPS-stimulated RAW 264.7 cells. The results are shown as relative expression against control expression without treatment. (e) The mRNA levels of M1 macrophages biomarkers were detected by real-time PCR. The results are shown as relative expression against control expression without treatment. (f) The secretion of proinflammatory cytokines were determined by ELISA. Data shown are the mean ± SD from 3 independent experiments. *P < 0.05, **P < 0.01 vs control group. #P < 0.05, ##P < 0.01 vs LPS-treated group. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/26725521), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Ang II increases the number of TSP-1 positive cells in mouse heart and skin.IHC staining of TSP-1 was performed on paraffin sections from the heart (A) and skin (B) of PBS and Ang II treated WT mice. Representative photographs are shown from five animals per group. C. TSP-1 protein levels were increased in Ang II treated HDMECs in a dose dependent manner (*p≤0.05). Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.pone.0109763), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Ba, Bv, Ca, Pm, Ye, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ba, Bv, Ca, Pm, Ye, Ze

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	NB600-1384AF350
Alexa Fluor 405	NB600-1384AF405
Alexa Fluor 488	NB600-1384AF488
Alexa Fluor 532	NB600-1384AF532
Alexa Fluor 594	NB600-1384AF594
Alexa Fluor 647	NB600-1384AF647
Alexa Fluor 700	NB600-1384AF700
Alexa Fluor 750	NB600-1384AF750
Allophycocyanin	NB600-1384APC
Biotin	NB600-1384B
DyLight 350	NB600-1384UV
DyLight 405	NB600-1384V
DyLight 488	NB600-1384G
DyLight 550	NB600-1384R
DyLight 594	NB600-1384DL594
DyLight 650	NB600-1384C
DyLight 680	NB600-1384FR
DyLight 755	NB600-1384IR
FITC	NB600-1384F
HRP	NB600-1384H
Janelia Fluor 549	NB600-1384JF549
Janelia Fluor 646	NB600-1384JF646
PE	NB600-1384PE
PerCP	NB600-1384PCP
mFluor Violet 450 SE	NB600-1384MFV450
mFluor Violet 500 SE	NB600-1384MFV500
mFluor Violet 610 SE	NB600-1384MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB600-1384SS

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NBP2-46892

LC3B Antibody (1251A) - BSA Free

(3 Reviews)

(12 Publications)

0.1 mg

NBP2-46892

Knockout Validated: LC3B Antibody (1251A) [NBP2-46892] - Western Blot analysis shows lysates of HeLa human cervical epithelial carcinoma parental cell line and LC3B knockout HeLa cell line (KO) untreated (-) or treated (+) with 50 uM Chloroquine for 18 hours. PVDF (Polyvinylidene difluoride) membrane was probed with 2.0 ug/mL of Rabbit Anti-Human/Mouse/Rat LC3B Monoclonal Antibody (1251A) [Catalog # NBP2-46892] followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog# HAF008). A specific band was detected for LC3B at a molecular weight of approximately 15 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. GAPDH is shown as a loading control. This experiment was conducted under reducing conditions.

<b>Genetic Strategies Validation. </b>Knockout Validated: LC3B Antibody (1251A) [NBP2-46892] - LC3B was detected in immersion fixed Chloroquine treated Hela cells (left) but was not detected in LC3B knockout Hela cells (right) using rabbit anti-human LC3B monoclonal antibody (1251A) [Catalog #NBP2-46892] at 0.3 ug/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm.

Western Blot: LC3B Antibody (1251A) [NBP2-46892] - The expression of LC3B in rat tissue. This image was submitted via customer Review.

<b>Biological Strategies Validation. </b>Flow (Intracellular): LC3B Antibody (1251A) [NBP2-46892] - Jurkat cells were either untreated (A) or treated with 50uM chloroquine for 24 hours (B). An intracellular stain was performed with anti-LC3B (1251A) antibody [Catalog # NBP2-46892] (blue) and a matched isotype control [Catalog # MAB1050] (orange). Cells were fixed with 4% paraformaldehyde, following fixation, cells were permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by rabbit IgG APC-conjugated secondary antibody (F0111, R&D Systems).

<b>Biological Strategies Validation. </b>Western Blot: LC3B Antibody (1251A) [NBP2-46892] - Effects of PDTC on LC3 on the hippocampus of rats harvested 12 h after CLP. Values of protein data are normalized to actin level and given in relative arbitrary units. LC3-II increased in CLP rats receiving vehicle or PDTC compared with those in sham-operated rats. PDTC + CLP rats showed an increase in LC3-II compared with vehicle-treated rats. Data were expressed as mean +/- SEM, n = 6/group. *P < 0.05 vs sham-operated group. # P < 0.05 vs 12 h or Veh + CLP group. PDTC pyrrolidine dithiocarbamate, LC-3 microtubule-associated protein light chain-3, CLP cecal ligation and puncture. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26067996) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: LC3B Antibody (1251A) [NBP2-46892] - HeLa cells were treated with Chloroquine for 24 hours prior to fixation, permeabilization and incubation with anti- [Catalog # NBP2-46892] and anti tubulin (NB100-690) antibodies. Image enlargement shows the accumulation of LC3 (green) on autophagosomes in response to Chloroquine treatment. Tubulin staining is shown in red and DNA is counterstained with DAPI (blue).

Immunohistochemistry-Paraffin: LC3B Antibody (1251A) [NBP2-46892] - IHC (Immunohistochemical) analysis of a formalin fixed and paraffin embedded tissue section of normal mouse brain using rabbit monoclonal LC3B (1251A) antibody [Catalog # NBP2-46892] at 1:100 dilution with HRP-DAB detection. The antibody generated a weak diffused cytoplasmic staining in most of the cells but some cells, especially within empty areas on the section, showed punctate signal also which signifies the presence of autophagy in those areas.

Flow Cytometry: LC3B Antibody (1251A) [NBP2-46892] - HeLa cells untreated or treated with 50 uM chloroquine for 24h. Image from the Alexa Fluor 405 version of this antibody.

<b>Biological Strategies Validation. </b>Western Blot: LC3B Antibody (1251A) [NBP2-46892] - Analysis shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with Chloroquine. PVDF membrane was probed with 0.2 ug/mL rabbit anti-LC3B monoclonal Antibody (1251A) (Catalog # NBP2-46892, Novus Biologicals), followed by 1:2000 dilution of goat anti-rabbit IgG secondary antibody. A specific band was detected for LC3B at a molecular weight of approximately 15 kDa in CQ treated NIH-3T3 and PC12 cell lines.

<b>Biological Strategies Validation. </b>Western Blot: LC3B Antibody (1251A) [NBP2-46892] - Western Blot analysis using HeLa cells treated with (+) or without (-) Chloriquine (CQ). A specific band was detected for LC3B at a molecular weight of approximately 15 kDa.

<b>Biological Strategies Validation. </b>Western Blot: LC3B Antibody (1251A) [NBP2-46892] - Western Blot image of monoclonal anti-LC3B Antibody (Clone 1251D) [Catalog # NBP2-46892]. HeLa and Neuro2A cells were treated with or without 50 uM chloroquine for 24 hours as indicated. Whole cell protein was then separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane for 30 min and blocked in 5% non-fat milk in TBST (Tris-buffered saline, 0.1% Tween 20). The membrane was probed with 2 ug/ml anti-LC3B Antibody in 1% milk, and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. Note the accumulation of LC3B II upon chloroquine treatment. Bands for LC3 were detected at a molecular weight of approximately 15 kDa in treated HeLA cells, and both treated and untreated Neuro2A cells.

Immunocytochemistry/Immunofluorescence: LC3B Antibody (1251A) [NBP2-46892] - HeLa cells were treated with 50 uM Chloroquine for 24 hour prior to fixation in 10% buffered formalin for 10 min. Cells were permeabilized in 0.1% Triton X-100 and incubated with 20 ug/ml anti- [Catalog # NBP2-46892] and 1:500 anti-tubulin [Catalog # NB100-690] for 1 h at room temperature. LC3B reactivity (green) was detected with ant-rabbit Dylight 488 and tubulin (red) with anti-mouse Dylight 550. Nuclei were counterstained with DAPI (blue).

Flow (Intracellular): LC3B Antibody (1251A) [NBP2-46892] - HeLa human cervical epithelial carcimoa cell line either treated with 50 uM chloroquine for 24 hours (filled histogram) or untreated (open histogram) was stained with Rabbit anti-Human LC3B Alexa Fluor 647 conjugated monoclonal antiobdy (1251A) [Catalog # NBP2-46892AF647]. To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation and Permeabilization Buffer Kit (FC012-NOV).

Immunoprecipitation: LC3B Antibody (1251A) [NBP2-46892] - Western blot analysis of LC3 immunoprecipitation. 10 ug of monoclonal anti-LC3B antibody (clone 1251A) [Catalog # NBP2-46892] was used to immunoprecipitate LC3 from 200 ug of total protein from HeLa cells treated with or without 50 uM chloroquine for 24 h. Antibody-protein was captured by magnetic protein A/G beads, washed and subjected to SDS-PAGE on a 4-15% gel. Protein was transferred to 0.2 um PVDF membrane and probed with 2 ug/ml anti-LC3B (NB100-2220) and detected with an anti-rabbit HRP conjugated secondary antibody using chemiluminescence. The detection of LC3 at a molecular weight of approximately 12 kDa and the antibody heavy chain (Ab Hc) at a molecular weight of approximately 50 kDa is indicated.

Reactivity:

Hu, Mu, Rt, Po, Bt, Bv, Fi, Xp, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bt, Bv, Fi, Xp, Ze

Applications:

, , , , ,

Applications: , , , , ,

Host:

Rabbit Monoclonal

Host: Rabbit Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP2-46892AF350
Alexa Fluor 405	NBP2-46892AF405
Alexa Fluor 488	NBP2-46892AF488
Alexa Fluor 532	NBP2-46892AF532
Alexa Fluor 594	NBP2-46892AF594
Alexa Fluor 647	NBP2-46892AF647
Alexa Fluor 700	NBP2-46892AF700
Alexa Fluor 750	NBP2-46892AF750
Allophycocyanin	NBP2-46892APC
Biotin	NBP2-46892B
DyLight 350	NBP2-46892UV
DyLight 405	NBP2-46892V
DyLight 488	NBP2-46892G
DyLight 550	NBP2-46892R
DyLight 594	NBP2-46892DL594
DyLight 650	NBP2-46892C
DyLight 680	NBP2-46892FR
DyLight 755	NBP2-46892IR
FITC	NBP2-46892F
HRP	NBP2-46892H
Janelia Fluor 549	NBP2-46892JF549
Janelia Fluor 646	NBP2-46892JF646
PE	NBP2-46892PE
PerCP	NBP2-46892PCP
mFluor Violet 450 SE	NBP2-46892MFV450
mFluor Violet 500 SE	NBP2-46892MFV500
mFluor Violet 610 SE	NBP2-46892MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP2-46892SS
Azide and BSA Free	NBP2-80829

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NBP2-59800

LC3B Antibody (1251D) - BSA Free

(1 Publication)

0.1 mg

NBP2-59800

Knockout Validated: LC3B Antibody (1251D) [NBP2-59800] - HeLa human cervical epithelial carcinoma parental cell line and LC3B knockout HeLa cell line (KO) untreated (-) or treated (+) with 50 uM Chloroquine for 18 hrs. PVDF membrane was probed with 0.1 ug/mL of Rabbit Anti-LC3B Monoclonal Antibody (1251D) [Catalog# NBP2-59800] followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog# HAF008). A specific band was detected for LC3B at approximately 15 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions.

<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Western Blot: LC3B Antibody (1251D) [NBP2-59800] - Total protein from HeLa and Neuro2A cells treated with or without 50 uM chloroquine for 24 hrs was separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF (Polyvinylidene difluoride) membrane and blocked in 5% non-fat milk in TBST (Tris-buffered saline, 0.1% Tween 20). The membrane was probed with 2.0 ug/ml anti-LC3B Antibody (1251d) [Catalog # NBP2-59800] in blocking buffer and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. Note the detection of LC3I and LC3II at a molecular weight of approximately 15 kDa.

Immunocytochemistry/Immunofluorescence: LC3B Antibody (1251D) [NBP2-59800] - HeLa cells were treated with 50uM CQ (Chloroquine) overnight, fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-LC3B (1251d) [Catalog # NBP2-59800] at 5 ug/ml overnight at 4C and detected with an anti-mouse DyLight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry-Paraffin: LC3B Antibody (1251D) [NBP2-59800] - Analysis of a FFPE (Formalin-Fixed Paraffin-Embedded) tissue section of mouse brain using anti-LC3B antibody (clone 1251D) [Catalog # NBP2-59800] at 5ug/ml concentration (1:200 dilution). The primary antibody binding to LC3 in cells was detected using HRP conjugated anti-Rabbit secondary antibody with DAB (3,3'Diaminobenzidine) reagent, and the sections were further counterstained with hematoxylin for labeling cellular nuclei. This LC3 antibody generated a diffused staining in all cell types (except the endothelial cells of blood vessels) and the signal was strongest in a subset of neuronal cells.

<b>Biological Strategies Validation. </b>Flow (Intracellular): LC3B Antibody (1251D) [NBP2-59800] - HeLa cells were either (A) untreated or (B) treated with 50uM chloroquine for 24 hrs. An intracellular stain was performed with anti-LC3B (1251D) antibody [Catalog # NBP2-59800] (blue) and a matched isotype control [Catalog # MAB1050] (orange). Cells were fixed with 4% paraformaldehyde, following fixation, cells were permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by rabbit IgG APC-conjugated secondary antibody (F0111, R&D Systems).

<b>Biological Strategies Validation. </b>Simple Western: LC3B Antibody (1251D) [NBP2-59800] - Lane view shows lysates of NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 50 uM Chloroquine for 18 hrs, loaded at 0.2 mg/ml. A specific band was detected for LC3B at approximately 17 kDa (as indicated) using 5 ug/mL of Rabbit Anti-Human LC3B Monoclonal Antibody (1251D) [Catalog# NBP2-59800]. This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , , , ,

Applications: , , , , ,

Host:

Rabbit Monoclonal

Host: Rabbit Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP2-59800AF350
Alexa Fluor 405	NBP2-59800AF405
Alexa Fluor 488	NBP2-59800AF488
Alexa Fluor 532	NBP2-59800AF532
Alexa Fluor 594	NBP2-59800AF594
Alexa Fluor 647	NBP2-59800AF647
Alexa Fluor 700	NBP2-59800AF700
Alexa Fluor 750	NBP2-59800AF750
Allophycocyanin	NBP2-59800APC
Biotin	NBP2-59800B
DyLight 350	NBP2-59800UV
DyLight 405	NBP2-59800V
DyLight 488	NBP2-59800G
DyLight 550	NBP2-59800R
DyLight 594	NBP2-59800DL594
DyLight 650	NBP2-59800C
DyLight 680	NBP2-59800FR
DyLight 755	NBP2-59800IR
FITC	NBP2-59800F
HRP	NBP2-59800H
Janelia Fluor 549	NBP2-59800JF549
Janelia Fluor 646	NBP2-59800JF646
PE	NBP2-59800PE
PerCP	NBP2-59800PCP
mFluor Violet 450 SE	NBP2-59800MFV450
mFluor Violet 500 SE	NBP2-59800MFV500
mFluor Violet 610 SE	NBP2-59800MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP2-59800SS
Azide and BSA Free	NBP2-80830

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NBP2-60735

LC3B Antibody (1251B) - BSA Free

0.1 mg

NBP2-60735

Knockout Validated: LC3B Antibody (1251B) [NBP2-60735] - Western blot analysis of HeLa human cervical epithelial carcinoma parental cell line and LC3B knockout HeLa cell line (KO) untreated (-) or treated (+) with 50uM Chloroquine for 18 hours. PVDF (polyvinylidene difluoride) membrane was probed with 0.1 ug/mL of anti-LC3B monoclonal antibody (Catalog # NBP2-60735) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for LC3B at approximately 15 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control.

<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Simple Western: LC3B Antibody (1251B) [NBP2-60735] - Lane view shows lysates of NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 50uM Chloroquine for 18 hours, loaded at 0.2 mg/mL. A specific band was detected for LC3B at approximately 17 kDa (as indicated) using 5 ug/mL of Rabbit Anti-Human/Mouse/Rat anti-LC3B Monoclonal Antibody (1251B) [Catalog # NBP2-60735]. This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Immunocytochemistry/Immunofluorescence: LC3B Antibody (1251B) [NBP2-60735] - HeLa cells were treated with 50uM CQ (Chloroquine) overnight, fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton X-100. The cells were incubated with anti- [Catalog # NBP2-60735] at 2 ug/ml overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

<b>Biological Strategies Validation. </b>Western Blot: LC3B Antibody (1251B) [NBP2-60735] - Total protein from HeLa and Neuro2A cells treated with or without 50 uM chloroquine (CQ) for 24 hours was separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF (Polyvinylidene difluoride) membrane and blocked in 5% non-fat milk in TBST (Tris-buffered saline, 0.1% Tween 20). The membrane was probed with 12.0 ug/ml anti-LC3B Antibody (1251B) [Catalog # NBP2-60735] in 1% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.

Immunohistochemistry-Paraffin: LC3B Antibody (1251B) [NBP2-60735] - Analysis of a FFPE (Formalin-Fixed Paraffin-Embedded) tissue section of mouse brain using anti-LC3B antibody (clone 1251B) [Catalog # NBP2-60735] at 5ug/ml concentration (1:200 dilution). The primary antibody binding to LC3 in cells was detected using HRP conjugated anti-Rabbit secondary antibody with DAB (3,3'Diaminobenzidine) reagent, and the sections were further counterstained with hematoxylin for labeling cellular nuclei. This LC3 antibody generated a diffused staining in all cell types (except the endothelial cells of blood vessels) and the signal was strongest in a subset of neuronal cells. A few cells depicted LC3 puncta formation/dotted staining in apparently autophagic cells.

<b>Biological Strategies Validation. </b>Flow (Intracellular): LC3B Antibody (1251B) [NBP2-60735] - HeLa cells were either (A) untreated or (B) treated with 50uM chloroquine for 24 hours. An intracellular stain was performed with anti-LC3B (1251B) antibody [Catalog # NBP2-60735] (blue) and a matched isotype control [Catalog # MAB1050] (orange). Cells were fixed with 4% paraformaldehyde, following fixation, cells were permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by rabbit IgG APC-conjugated secondary antibody (F0111, R&D Systems).

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , , , ,

Applications: , , , , ,

Host:

Rabbit Monoclonal

Host: Rabbit Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP2-60735AF350
Alexa Fluor 405	NBP2-60735AF405
Alexa Fluor 488	NBP2-60735AF488
Alexa Fluor 532	NBP2-60735AF532
Alexa Fluor 594	NBP2-60735AF594
Alexa Fluor 647	NBP2-60735AF647
Alexa Fluor 700	NBP2-60735AF700
Alexa Fluor 750	NBP2-60735AF750
Allophycocyanin	NBP2-60735APC
Biotin	NBP2-60735B
DyLight 350	NBP2-60735UV
DyLight 405	NBP2-60735V
DyLight 488	NBP2-60735G
DyLight 550	NBP2-60735R
DyLight 594	NBP2-60735DL594
DyLight 650	NBP2-60735C
DyLight 680	NBP2-60735FR
DyLight 755	NBP2-60735IR
FITC	NBP2-60735F
HRP	NBP2-60735H
Janelia Fluor 549	NBP2-60735JF549
Janelia Fluor 646	NBP2-60735JF646
PE	NBP2-60735PE
PerCP	NBP2-60735PCP
mFluor Violet 450 SE	NBP2-60735MFV450
mFluor Violet 500 SE	NBP2-60735MFV500
mFluor Violet 610 SE	NBP2-60735MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP2-60735SS
Azide and BSA Free	NBP2-80831

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NBP3-26154

LC3B Antibody (7D11)

100 ul

NBP3-26154

Western Blot: LC3B Antibody (7D11) [NBP3-26154] - Positive Western Blot detected in: HepG2 whole cell lysate, U87 whole cell lysate, RAW264.7 whole cell lysate, U251 whole cell lysate, Hela whole cell lysate. All lanes: LC3B Antibody at 1: 500 Secondary: Goat polyclonal to rabbit IgG at 1/50000 dilution. Predicted band size: 15 kDa Observed band size: 15 kDa

Flow Cytometry: LC3B Antibody (7D11) [NBP3-26154] - Overlay Peak curve showing Hela cells stained with LC3B Antibody (7D11) (red line) at 1:50. The cells were fixed in 4% formaldehyde and permeated by 0.2% Triton X-100. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/1*10^6 cells) for 45min at 4C. The secondary antibody used was FITC-conjugated Goat Anti-rabbit IgG (H+L) at 1:200 dilution for 35min at 4C. Control antibody (green line) was rabbit IgG (1ug/1*10^6 cells) used under the same conditions. Acquisition of >10,000 events was performed.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

, ,

Applications: , ,

Host:

Rabbit Monoclonal

Host: Rabbit Monoclonal

Conjugate:

Unconjugated

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NB600-1384C

LC3B Antibody [DyLight 650]

0.1 ml

Reactivity:

Hu, Mu, Rt, Po, Ba, Bv, Ca, Pm, Ye, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ba, Bv, Ca, Pm, Ye, Ze

Applications:

, , , , , , , , , ,

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

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

DyLight 650

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB600-1384AF350
Alexa Fluor 405	NB600-1384AF405
Alexa Fluor 488	NB600-1384AF488
Alexa Fluor 532	NB600-1384AF532
Alexa Fluor 594	NB600-1384AF594
Alexa Fluor 647	NB600-1384AF647
Alexa Fluor 700	NB600-1384AF700
Alexa Fluor 750	NB600-1384AF750
Allophycocyanin	NB600-1384APC
Biotin	NB600-1384B
DyLight 350	NB600-1384UV
DyLight 405	NB600-1384V
DyLight 488	NB600-1384G
DyLight 550	NB600-1384R
DyLight 594	NB600-1384DL594
DyLight 680	NB600-1384FR
DyLight 755	NB600-1384IR
FITC	NB600-1384F
HRP	NB600-1384H
Janelia Fluor 549	NB600-1384JF549
Janelia Fluor 646	NB600-1384JF646
PE	NB600-1384PE
PerCP	NB600-1384PCP
mFluor Violet 450 SE	NB600-1384MFV450
mFluor Violet 500 SE	NB600-1384MFV500
mFluor Violet 610 SE	NB600-1384MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB600-1384SS
BSA Free	NB600-1384

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