Product Search for: Novus Biologicals

H00008878-M01

p62/SQSTM1 Antibody (2C11) - Azide and BSA Free

(9 Reviews)

(441 Publications)

0.1 mg

H00008878-M01

Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - ICC-IF analysis of SQSTM1 protein in HeLa cell using SQSTM1/p62 antibody (clone 2C11) at 10 ug/ml concentration.

Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Analysis of p62 in SH-SY5Y cells using anti-p62/SQSTM1 antibody. Red - p62 puncta; Blue - nuclear DAPI; Green - Cytoskeleton. Image from verified customer review.

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Disruption of the VANGL2-p62/SQSTM1 interaction in breast cancer cells. Soft agar colony formation of T47D cells overexpressing GFP, GFP-VANGL2 and GFP-p62/SQSTM1 (right). Protein expression was revealed with anti-p62/SQSTM1, anti-VANGL2 and anti-GFP antibodies by western blot analysis (right). In anti-GFP blot, the arrowhead indicates position of co-migrating GFP-VANGL2 and GFP-p62/SQSTM1 and the asterisk pinpoints GFP alone. Error bars represent mean+/-s.d. (n=3). Image collected and cropped by CiteAb from the following publication (//www.nature.com/doifinder/10.1038/ncomms10318), licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Analysis of p62/SQSTM1 antibody on HeLa cells. Image from verified customer review.

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Confirmation of loss of hepatic ATG7 protein and a phenotype of hepatic autophagy inactivation. Fourth ug of liver protein were loaded on 4-20% gradient Tris-Glycine gels and subsequently immunoblotted with antibodies against ATG7, p62 and LC3. Blots were visualized with G:BOX Chemi XRQ (SynGene). Representative immunoblots are shown for WT and KO mice. Full-length immunoblots are presented in Supplementary Figure S1. ATG7: autophagy-related protein 7; p62: sequestosome 1; LC3: microtubule-associated protein 1 light chain 3; WT: Mx1-Cre-Atg7F/F mouse; KO: Mx1-Cre+Atg7F/F mouse. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-14405-w), licensed under a CC-BY license.

ELISA: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Detection limit for recombinant GST tagged SQSTM1 is approximately 0.03ng/ml as a capture antibody.

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Activation of multiple signaling pathways by dandelion root extractA. Western blots of proteins involved in programmed cell death & cell survival & inflammation B. Densitometry quantification of western blot analysis from three independent experiments. *P<0.05, **P< 0.001. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.11485), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Elevated levels of additional proteins & GM3 ganglioside in the MEC of MPS IIIB brain.Staining performed with antibodies to the indicated substances was observed in the MEC region of 3 month-old MPS IIIB mice (for total ubiquitin & polyubiquitin) & 6 months for all others. Staining was not seen in the MEC region of age-matched control mice (Naglu +/−) nor in the LEC region of MPS IIIB mice (the latter not shown). Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0027461), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Confirmation of loss of hepatic ATG7 protein & a phenotype of hepatic autophagy inactivation. Fourty µg of liver protein were loaded on 4–20% gradient Tris-Glycine gels & subsequently immunoblotted with antibodies against ATG7, p62 & LC3. Blots were visualised with G:BOX Chemi XRQ (SynGene). Representative immunoblots are shown for WT & KO mice. Full-length immunoblots are presented in Supplementary Figure S1. ATG7: autophagy-related protein 7; p62: sequestosome 1; LC3: microtubule-associated protein 1 light chain 3; WT: Mx1-Cre−Atg7F/F mouse; KO: Mx1-Cre+Atg7F/F mouse. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29074879), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - The content of mitochondrial p62, OPTN, & VCP in spinal cord of SOD1‐G93A mice is decreased by Parkin knockoutImmunostaining of anterior horn of lumbar spinal cord for ChAT (in green) & p62 (in red) at 130 days. Scale bar, 10 μm.Quantification of the percentage of ChAT‐positive motor neurons (MN) containing p62‐positive inclusions at 130 days. PKO/G93A MN have less SOD1 aggregates than G93A, while Non Tg & PKO MN have none. Data were collected from n = 4 (two males & two females) mice for Non Tg & PKO & n = 6 (three males & three females) for G93A & PKO/G93A. Number of images recorded for each genotype: n = 9 for Non Tg & PKO, n = 16 for G93A, & n = 12 for PKO/G93A. Results are expressed as mean ± SEM; *P = 0.032, by unpaired one‐tailed Student's t‐test.Representative Western blots of OPTN, p62, & VCP in spinal cord mitochondrial fractions at 130 days. Protein levels are normalized by Complex V. The quantifications in panels (D–F) show that mitochondria of PKO/G93A mice have less mitophagy adaptor proteins than G93A relative to Complex V.Quantification of OPTN relative to Complex V at 130 days. Results are expressed as mean ± SEM & as percent of Non Tg; n = 8 (four males & four females); *P = 0.039 by paired Student's t‐test.Quantification of p62, relative to Complex V at 130 days. Results are expressed as mean ± SEM & as percent of Non Tg; n = 8 (four males & four females); **P = 0.0029 by paired Student's t‐test.VCP was quantified using Complex V as loading reference at 130 days. Results are expressed as mean ± SEM & as percent of Non Tg; n = 8 (four males & four females); *P = 0.044 by paired Student's t‐test.Source data are available online for this figure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30126943), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - miR-29a downregulates TFEB & ATG9A to inhibit autophagyA. Schematic representation of the miR-29 family members & 3′-UTR binding sites of miR-29 targets as well as mutated binding sites used in Luciferase Assays: Transcription Factor EB (TFEB) & Autophagy-related protein 9A (ATG9A). All three miR-29 family members (miR-29a, miR-29b, & miR-29c) have identical seed sequences. Conserved miR-29 binding sites in the 3′-UTR of mRNA transcripts encoding ATG9A & TFEB are depicted in bold. B. 10ug of total protein cell lysates from Panc-1 transfected with CTRL or miR-29a mimics were subjected to western blot analysis for TFEB, ATG9A, & GAPDH. Relative quantification of band intensities normalized to GAPDH are shown below respective blots. C. Relative firefly luciferase activity from TFEB & ATG9A 3′ UTR wild type (WT) & mutant (mut) reporter constructs following co-transfection into Panc-1 cells with control or miR-29a mimics. All readouts were normalized to renilla luciferase activity for each well. Average relative luminesce normalized to respective controls is presented (n=6) ± S.E.M. D. 5ug of total protein cell lysates from Panc-1 cells were transfected with CTRL, miR-29a mimics, siCTRL, siTFEB, or siATG9A. 24 hours post-transfection, total protein was harvested & subjected to western blot analysis for p62 & LC3B, & GAPDH was used as loading control. Quantification of band intensities normalized to GAPDH & relative to control are shown below respective blots. All experiments were repeated 3 times & representative data is presented. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.11928), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Effects of autophagy inhibitors in combination with IFN-gamma on expression of PD-L1 in gastric cancer cell lines. a The effect of chloroquine (CQ) or 3-MA on expression of PD-L1 with or without INF-gamma for 24 h was determined by flow cytometry assays. In AGS & NCI-n87 cells, MFI as the indication of PD-L1 expression level can be further increased by the treatment of INF-gamma . b Levels of LC3B-I/II, p62/SQSTM1 & PD-L1 were determined by Western blots in AGS & NCI-n87 cells treated by CQ, 3-MA, bafilomycin A1 (Baf) or rapamycin (Rap) for 24 h. c Positive staining of PD-L1 (red) & LC3 positive puncta (green) was determined by immunofluorescence in AGS & NCI-n87 cells treated by autophagy inhibitors & activator as in (b). d Rapamycin decreased the levels of PD-L1 protein in AGS & NCI-n87 cells as shown by flow cytometry. Results were averaged & blots were representative of 4 independent experiments, *p < 0.05, **p < 0.01 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30925913), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Immunocytochemistry/ Immunofluorescence: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - The content of mitochondrial p62, OPTN, & VCP in spinal cord of SOD1‐G93A mice is decreased by Parkin knockoutImmunostaining of anterior horn of lumbar spinal cord for ChAT (in green) & p62 (in red) at 130 days. Scale bar, 10 μm.Quantification of the percentage of ChAT‐positive motor neurons (MN) containing p62‐positive inclusions at 130 days. PKO/G93A MN have less SOD1 aggregates than G93A, while Non Tg & PKO MN have none. Data were collected from n = 4 (two males & two females) mice for Non Tg & PKO & n = 6 (three males & three females) for G93A & PKO/G93A. Number of images recorded for each genotype: n = 9 for Non Tg & PKO, n = 16 for G93A, & n = 12 for PKO/G93A. Results are expressed as mean ± SEM; *P = 0.032, by unpaired one‐tailed Student's t‐test.Representative Western blots of OPTN, p62, & VCP in spinal cord mitochondrial fractions at 130 days. Protein levels are normalized by Complex V. The quantifications in panels (D–F) show that mitochondria of PKO/G93A mice have less mitophagy adaptor proteins than G93A relative to Complex V.Quantification of OPTN relative to Complex V at 130 days. Results are expressed as mean ± SEM & as percent of Non Tg; n = 8 (four males & four females); *P = 0.039 by paired Student's t‐test.Quantification of p62, relative to Complex V at 130 days. Results are expressed as mean ± SEM & as percent of Non Tg; n = 8 (four males & four females); **P = 0.0029 by paired Student's t‐test.VCP was quantified using Complex V as loading reference at 130 days. Results are expressed as mean ± SEM & as percent of Non Tg; n = 8 (four males & four females); *P = 0.044 by paired Student's t‐test.Source data are available online for this figure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30126943), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

$Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - G2019S LRRK2 hippocampal neurons do not have elevated alpha -synuclein pathology 14 days post-transduction. a Primary hippocampal neurons from NTG or G2019S pups were transduced with 2.5 μg/mL alpha -synuclein PFFs & allowed to age a further 14 days prior to sequential detergent fractionation. TX-100-insoluble alpha -synuclein & p62 are similar in both neuron types. b Quantification of soluble proteins shows ~ 25-fold elevation in the expression of LRRK2 & a commensurate ~ 50-fold elevation in pS395 LRRK2, indicative of the elevated LRRK2 kinase activity associated with the G2019S mutation. Soluble alpha -synuclein levels were equivalent between the cultures. c No significant differences were found between the genotypes in insoluble proteins by an unpaired t-test with Welch’s correction. (N = 3 biological replicates for each protein). Means + s.e.m.; **P < 0.01; *P < 0.05 by an unpaired t-test with Welch’s correction for unequal variances. All values are normalized to NTG neurons treated with alpha -synuclein PFFs & DMSO Image collected & cropped by CiteAb from the following publication (//actaneurocomms.biomedcentral.com/articles/10.1186/s40478-018-0550-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

$Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - LRRK2 inhibition does not reduce insoluble alpha -synuclein in wildtype hippocampal neurons. a Primary hippocampal neurons from CD1 pups were treated with 30 nM LRRK2 inhibitors PF-475, PF-360 or DMSO as a vehicle control, then transduced with 2.5 μg/mL alpha -synuclein PFFs & allowed to age a further 14 days prior to sequential detergent fractionation. TX-100-insoluble alpha -synuclein & p62 are similar in both LRRK2 inhibitor treated & untreated neurons. b Quantification of soluble proteins show some reduction of LRRK2 protein levels PF-475 treatment & ~ 75% inhibition of LRRK2 activity (as assayed by pS935 LRRK2) by both PF-475 & PF-360. c Insoluble pS129 alpha -synuclein was slightly, but significantly elevated by PF-360 treatment, while alpha -synuclein & p62 were unchanged by one-way ANOVA. (N = 5 biological replicates for each protein). Means + s.e.m.; *P < 0.05, ****p < 0.0001 by one-way ANOVA with Dunnett’s multiple comparison test. All values are normalized first to GAPDH, as a loading control, then to DMSO-treated neurons Image collected & cropped by CiteAb from the following publication (//actaneurocomms.biomedcentral.com/articles/10.1186/s40478-018-0550-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Immunocytochemistry/ Immunofluorescence: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Colocalization of VANGL2 & p62/SQSTM1 in late endosomes.(a) Immunofluorescence staining of SKBR7 cells showed colocalization of endogenous VANGL2 (green) & p62/SQSTM1 (red) in discrete cytoplasmic puncta. Scale bar, 10 μm. The mean Pearson correlation for VANGL2 & p62/SQSTM1 is 0.62, calculated using the Image J software for ∼15 cells per field of view & from 10 images. (b) Partial colocalization of VANGL2 & p62/SQSTM1 in late endosomes of SKBR7 cells stained w/ the LAMP1 marker. Scale bar, 20 μm. (c) SKBR7 ells cultured in a nutrient-deprived medium & treated w/ 100 nM bafilomycin A1 (6 h) before fixation. Double labelling against VANGL2 (arrowheads) & p62/SQSTM1 (arrows), as described in Methods, showed accumulations of both markers in vesicular structures probably resembling endosomes/amphisomes. Scale bar, 200 nm. (d) IMCD3 cells treated w/ PBS or EGTA (5 mM) for 30 min. Immunofluorescence & confocal analysis performed using the indicated antibodies. Scale bar, 10 μm. Inserts show colocalized VANGL2 & p62/SQSTM1. (e) The VANGL2–p62/SQSTM1 complex recovered in confluent SKBR7 or IMCD3 cells w/ 2G4 mAb (IP VANGL2) but not a control antibody (IP IgG2a ctrl) as seen using western blot analysis w/ the indicated antibodies. The complex more abundant in cancer cells (SKBR7) than in polarized cells (IMCD3). Note that human (SKBR7 cells) & murine (IMCD3 cells) p62/SQSTM1 run at different molecular weights. (f) Lysates of confluent IMCD3 cells treated for 30 min w/ PBS or EGTA subjected to 2G4 mAb immunoprecipitation (IP VANGL2). Immunoprecipitated proteins probed by western blot analysis w/ the indicated antibodies. Increased amounts of VANGL2–p62/SQSTM1 complexes recovered after EGTA treatment. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms10318), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Colocalization of VANGL2 & p62/SQSTM1 in late endosomes.(a) Immunofluorescence staining of SKBR7 cells showed colocalization of endogenous VANGL2 (green) & p62/SQSTM1 (red) in discrete cytoplasmic puncta. Scale bar, 10 μm. The mean Pearson correlation for VANGL2 & p62/SQSTM1 is 0.62, calculated using the Image J software for ∼15 cells per field of view & from 10 images. (b) Partial colocalization of VANGL2 & p62/SQSTM1 in late endosomes of SKBR7 cells stained w/ the LAMP1 marker. Scale bar, 20 μm. (c) SKBR7 ells cultured in a nutrient-deprived medium & treated w/ 100 nM bafilomycin A1 (6 h) before fixation. Double labelling against VANGL2 (arrowheads) & p62/SQSTM1 (arrows), as described in Methods, showed accumulations of both markers in vesicular structures probably resembling endosomes/amphisomes. Scale bar, 200 nm. (d) IMCD3 cells treated w/ PBS or EGTA (5 mM) for 30 min. Immunofluorescence & confocal analysis performed using the indicated antibodies. Scale bar, 10 μm. Inserts show colocalized VANGL2 & p62/SQSTM1. (e) The VANGL2–p62/SQSTM1 complex recovered in confluent SKBR7 or IMCD3 cells w/ 2G4 mAb (IP VANGL2) but not a control antibody (IP IgG2a ctrl) as seen using western blot analysis w/ the indicated antibodies. The complex more abundant in cancer cells (SKBR7) than in polarized cells (IMCD3). Note that human (SKBR7 cells) & murine (IMCD3 cells) p62/SQSTM1 run at different molecular weights. (f) Lysates of confluent IMCD3 cells treated for 30 min w/ PBS or EGTA subjected to 2G4 mAb immunoprecipitation (IP VANGL2). Immunoprecipitated proteins probed by western blot analysis w/ the indicated antibodies. Increased amounts of VANGL2–p62/SQSTM1 complexes recovered after EGTA treatment. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms10318), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Parkin protein levels are unaffected in liver while a small decrease is measured in G93A cerebellumRepresentative Western blot for Parkin & p62 in cerebellum (Cb) homogenates at 130 days.Quantification indicates that Parkin is decreased in G93A mice relative to Non Tg at 130 days. Protein levels were normalized by beta ‐actin. Results are expressed as mean ± SEM & percent of Non Tg; n = 8 (four males & four females) mice per group. *P = 0.039 by paired Wilcoxon's test.Quantification of p62 in the homogenates indicates that p62 is increased in PKO & PKO/G93A mice, independent of SOD1‐G93A expression, at 130 days of age. beta ‐actin was used for normalization. Results are expressed as mean ± SEM & percent of Non Tg; n = 8 (four males & four females) mice per group. No statistically significant differences were found between Non Tg & G93A by paired Friedman's test with Dunn's correction (P = 0.99). ***P = 0.0001 by paired Student's t‐test (G93A vs. PKO/G93A).Representative Western blot of Parkin & p62 in liver homogenates at 130 days.Parkin protein levels were quantified at 130 days. beta ‐actin was used for normalization. Results are expressed as mean ± SEM & percent of Non Tg; n = 8 (four males & four females) mice per group. No statistically significant differences were found between Non Tg & G93A (P = 0.546 by paired Wilcoxon's test).Quantification of p62 protein levels in liver at 130 days. Results are expressed as mean ± SEM & percent of Non Tg; n = 8 (four males & four females) mice per group. No statistically significant differences were found between Non Tg & G93A (P = 0.546 by paired Wilcoxon's test). No statistically significant differences were found among the other groups.Source data are available online for this figure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30126943), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Parkin knockout mitigates the accumulation of mitophagy adaptors in G93A mitochondria at disease end stageA, BRepresentative Western blots of p62 (A), OPTN, & VCP (B) in spinal cord mitochondria from end‐stage mice.Cp62 quantification, using Complex V as normalizer, shows a strong accumulation of p62 in SOD1‐G93A mitochondria at end stage. Results are expressed as mean ± SEM & as percent of Non Tg; n = 8 (four males & four females) mice per group. No statistically significant differences were found between G93A & PKO/G93A (P = 0.078 by paired Wilcoxon's test); ***P = 0.0007 (Non Tg vs. G93A) & *P = 0.037 (PKO & PKO/G93A) both by paired Friedman's test with Dunn's correction.DQuantification of OPTN accumulation in end‐stage mitochondria, with Complex V as protein loading control. Results are expressed as mean ± SEM & as percent of Non Tg; n = 8 (four males & four females) mice per group. **P = 0.0078 (for G93A & PKO/G93A) by paired Wilcoxon's test; ***P = 0.0003 (Non Tg vs. G93A) by paired Friedman's test with Dunn's correction. No other statistically significant differences were found.EQuantification of VCP in mitochondria at disease end stage. Complex V was used as normalizer. Results are expressed as mean ± SEM & as percent of Non Tg; n = 8 (four males & four females) mice per group. *P = 0.039 (for G93A & PKO/G93A) by paired Wilcoxon's test; ***P = 0.0007 (Non Tg vs. G93A) & *P = 0.037 (PKO & PKO/G93A) both by paired Friedman's test with Dunn's correction.Source data are available online for this figure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30126943), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Bv, Ha, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, Bv, Ha, Rb

Applications:

, , , ,

Applications: , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

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Unconjugated

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H00023435-M01

TDP-43/TARDBP Antibody (2E2-D3) - Azide and BSA Free

(3 Reviews)

(212 Publications)

0.1 mg

H00023435-M01

Western Blot: TDP-43/TARDBP Antibody (2E2-D3) [H00023435-M01] - PDI co-precipitates with TDP-43/TARDBP and is increased in mutant TDP-43 transgenic mouse spinal cords. Co-immunoprecipitation of wildtype and mutant Q331K TDP-43-mCherry from Neuro2a cell lysates using an anti-PDI antibody followed by TDP-43 immunoblot. Control immunoprecipitations were performed using untransfected (Untrans) or mCherry expressing cell lysates, buffer only with coprecipitating antibodies or TDP-43 Q331K cell lysates using an irrelevant, isotype-matched control antibody (IgG). Input control (2%) shows expression of mCherry-TDP43 in wildtype and Q331K cell lysates. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0081170) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: TDP-43/TARDBP Antibody (2E2-D3) [H00023435-M01] - Analysis of monoclonal antibody to TARDBP on HeLa cell. Antibody concentration 10 ug/ml.

Immunohistochemistry-Paraffin: TDP-43/TARDBP Antibody (2E2-D3) [H00023435-M01] - TARDBP on formalin-fixed paraffin-embedded human leiomyosarcoma. [antibody concentration 3 ug/ml]

Western Blot: TDP-43/TARDBP Antibody (2E2-D3) [H00023435-M01] - Analysis of TARDBP expression in transfected 293T cell line by TARDBP monoclonal antibody (M01), clone 2E2-D3.Lane 1: TARDBP transfected lysate(44.7 KDa).Lane 2: Non-transfected lysate.

Immunohistochemistry-Paraffin: TDP-43/TARDBP Antibody (2E2-D3) [H00023435-M01] - Analysis of monoclonal antibody to TARDBP on formalin-fixed paraffin-embedded human leiomyosarcoma tissue Antibody concentration 3 ug/ml.

<b>Genetic Strategies Validation. </b>Western Blot: TDP-43/TARDBP Antibody (2E2-D3) [H00023435-M01] - Analysis of TARDBP over-expressed 293 cell line, cotransfected with TARDBP Validated Chimera RNAi ( Cat # H00023435-R01V ) (Lane 2) or non-transfected control (Lane 1). Blot probed with TARDBP monoclonal antibody (M01), clone 2E2-D3 (Cat # H00023435-M01 ). GAPDH ( 36.1 kDa ) used as specificity and loading control.

ELISA: TDP-43/TARDBP Antibody (2E2-D3) [H00023435-M01] - Detection limit for recombinant GST tagged TARDBP is 0.3 ng/ml as a capture antibody.

Western Blot: TDP-43/TARDBP Antibody (2E2-D3) [H00023435-M01] - PDI co-precipitates with TDP-43 & is increased in mutant TDP-43 transgenic mouse spinal cords.(A) Co-immunoprecipitation of wildtype & mutant Q331K TDP-43-mCherry from Neuro2a cell lysates using an anti-PDI antibody followed by TDP-43 immunoblot. Control immunoprecipitations performed using untransfected (Untrans) or mCherry expressing cell lysates, buffer only with coprecipitating antibodies or TDP-43 Q331K cell lysates using an irrelevant, isotype-matched control antibody (IgG). Input control (2%) shows expression of mCherry-TDP43 in wildtype & Q331K cell lysates. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0081170), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TDP-43/TARDBP Antibody (2E2-D3) [H00023435-M01] - CLIP shows that TDP-43 directly interacts with translational target mRNAs. (A) Schematic diagram of the CLIP experiment. (B) Immunoblot showing specific IP of TDP-43 from MN1 cells under CLIP conditions. A TDP-43 band is specifically observed in the input & TDP-43 IP lanes from both untreated & UV-treated cells, but not in the IgG or anti-RBP X control lanes. RBP X is an IP of a different RNA-binding protein, which serves as an additional specificity control here. (C and D) Plots showing percentage of input mRNA in the TDP-43 IP for the mRNAs indicated in three separate CLIP assays from MN1 cells. Performing CLIP from UV-treated cells leads to significant recovery & enrichment relative to Gapdh for all mRNAs examined (C). In the absence of UV treatment, much less mRNA is recovered & no significant enrichment relative to Gapdh mRNA is observed. Note that the Y-axis scale in D is 1000× smaller than in C, indicating strong crosslinking dependence of mRNA co-immunoprecipitation (n = 3, s.e.m. error bars, P-values indicated in the plot, unpaired two-tailed t-test). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30357366), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: TDP-43/TARDBP Antibody (2E2-D3) [H00023435-M01] - SINE compounds partially rescue motor deficits in an animal model of ALS/FTD. (a) Schematic timeline for in vivo experiments. (b) Western blot demonstrating the expression of human TDP43 within the brains of AAV9-injected animals, which we have previously shown is largely restricted to neuronal cell types (Jackson et al.59). Full-length blots appear in Fig. S1. (c) The hang test measures the time it takes for animals to fall from grid-like scaffold when it is lifted & tilted. A single representative animal administered AAV9-TDP43 is shown; note the inability of the hind limbs to clasp the wire mesh. (d) TDP43 expression impaired motor performance on the hang test at 3 weeks, an effect that could be mitigated by 7.5 mg kg-1 KPT-350. (e) Rotarod testing at 4 weeks of age showed prominent deficits associated with TDP43 expression that were resistant to treatment with SINE compounds. n, number of animals per condition. ns, not significant, mean ± s.e.m. *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA with Bonferroni correction. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-018-22858-w), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Ba, I, Pm, Pp

Species Glossary

Reactivity: Hu, Mu, Rt, Ba, I, Pm, Pp

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Conjugate:

Unconjugated

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NB100-384

Histone H2AX [p Ser139] Antibody - BSA Free

(1 Review)

(180 Publications)

0.1 ml

NB100-384

Detection of Human and Mouse Histone H2AX [p Ser139] by Western Blot. Samples: Nuclear extract (50 ug) from human HEK293, human melanoma (G361), mouse wildtype embryonic fibroblasts (+/+) or mouse H2AX knockout embryonic fibroblasts (-/-). Antibody: Affinity purified rabbit Histone H2AX [p Ser139] antibody NB100-384 used at 0.1 ug/ml. Detection: Chemiluminescence with 30 second exposure. (NCS, neocarzinostatin - 200 ng/ml, 30 min). Bands appear at an observed molecular weight of ~15 kDa.

<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Samples: Neocarzinostatin treated asynchronous HeLa cells (left) and untreated asynchronous HeLa cells (right) . Antibody: Affinity purified rabbit Histone H2AX [p Ser139] used at a dilution of 1:5,000 (0.2ug/ml). Detection: Red fluorescent Anti-rabbit IgG-DyLight 594 used at a dilution of 1:100.

Simple Western lane view shows a specific band for Histone H2AX [p Ser139] in 0.2 mg/ml of Jurkat lysate(s). This experiment was performed under reducing conditions using the 12 - 230 kDa separation system.

<b>Biological Strategies Validation. </b>Immunocompromised mice were subcutaneously injected with cancerous cell lines and tumors were allowed to establish. Treatments occurred every other day and the studied compound or the equivalent vehicle control administered intraperitoneally for five weeks. Tumor volume and mass were measured two times per week. IHC analysis of sectioned tumor tissues from the MDA-MB-231 study. Each section was subjected to the specified antibody followed by a biotinylated secondary antibody. Detection was done using a DAB Peroxidase HRP Substrate Kit (brown) followed by Hematoxylin counterstaining (purple). Images were obtained using inverted bright field microscopy. Sectioning results are representative of three individual tumors. Scale bar is 50 microns. Image collected and cropped by CiteAb from the following publication (nature.com/articles/s41598-017-01230-4), licensed under a CC-BY license.

Immunostaining of Histone H2AX [p Ser139] WT1 and 5mC in patients with IgA nephropathy and controls. Examples of PAS staining and immunostaining with Histone H2AX [p Ser139] (green) and WT1 (red), pATM and 5mC in glomeruli of IgA nephropathy and controls. A kidney sample of a 65-year-old male of IgA nephropathy without podocytopathic features. Arrows indicate Histone H2AX [p Ser139] and WT1 double-positive cells. Scale bars: 50um. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-019-57140-0) licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>Samples: Nuclear extract from HeLa cells treated with 100 uM EPE for 4 hours (+) or mock treated (-). Antibody: Affinity purified rabbit Histone H2AX [p Ser139] antibody used at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 3 minutes. Band appears at an observed molecular weight of ~17 kDa.

<b>Biological Strategies Validation. </b>Detection of human Histone H2AX [p Ser139] by western blot. Samples: Whole cell lysate (50 ug) from Jurkat cells treated with 100 uM EPE for 4 hours (+) or mock treated (-). Antibody: Affinity purified rabbit Histone H2AX [p Ser139] antibody NB100-384 used for WB at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 3 seconds. Band appears at an observed molecular weight of ~18 kDa.

FFPE section of mouse CT26 colon carcinoma. Antibody: Affinity purified rabbi Histone H2AX [p Ser139] antibody used at a dilution of 1:1,000 (1 ug/ml). Detection: DAB.

Analysis of Histone H2AX [p Ser139] in EPE Treated Jurkat Cells. Cells were treated for 3 hrs in 5ug/ml etoposide, fixed in 1.5% PFA, and permeabilized in 90% Methanol. 1 million cells were stained with 0.5 ug anti-KLH or anti-H2AX NB100-384 and secondary FITC-conjugated goat anti-rabbit (in a 150ul reaction). Black- etosposide treated, anti-KLH; Red- untreated, anti-Histone H2AX [p Ser139]; Blue- etoposide treated , anti-Histone H2AX [p Ser139].

FFPE section of human ovarian carcinoma. Antibody: Affinity purified rabbit Histone H2AX [p Ser139] antibody used at a dilution of 1:5,000 (0.2 ug/ml). Detection: DAB.

Electropherogram image(s) of corresponding Simple Western lane view. Histone H2AX [p Ser139] antibody was used at 5 ug/ml dilution on Jurkat lysate(s).

Immunohistochemistry: Rabbit Polyclonal Histone H2AX [p Ser139] Antibody - Histone H2AX Antibody on mouse cancer tissue. H2AX (Gray) and H2BGFP(Green). Primary antibody dilution: 1:1000 in a 10um slice.

mTORC1/2 activity prevents Cisplatin-induced cell death in MCF-10A cells. (A) Western blot displaying effects on mTOR signaling during a dose escalation of PP242 treatment in MCF-10A cells; (B) Western blot displaying effects of mTOR signaling on a dose escalation of cisplatin treatment in MCF-10A cells; (C) Western blot displaying effects on mTOR signaling and cell death during non-treated, Cisplatin, PP242, and Cisplatin + PP242-treated MCF-10A cells.

Western Blot: Histone H2AX [p Ser139] Antibody [NB100-384] - ATM & NF-kappa B activation are downregulated in Ercc1-/ delta mice heterozygous for Atm. (A) Livers were collected at 12 weeks of age from WT, Ercc1-/ delta & Ercc1-/ delta Atm+/- mic (n=3 per genotype) & lysates analyzed by western blot for activation of ATM & its downstream effectors. (B) Same liver lysates were used to measure phosphorylation of p65 & I kappa B alpha . (C) Western blot analysis of livers from 16-week-old WT, Ercc1-/ delta & Ercc1-/ delta Atm+/-mice (n=3 per genotype) probed for activation of ATM. GAPDH was used as a loading control. (D) Same liver lysates used to measure activation of NF-kappa B. (E) Fourteen-week-old livers from Ercc1-/ delta & Ercc1-/ delta p65+/- mice (n=3 per genotype) were analyzed by western blot for activation of ATM (F) & NF-kappa B. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32201398), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Histone H2AX [p Ser139] Antibody [NB100-384] - Immunostaining of gamma H2AX, WT1 & 5mC in patients with IgA nephropathy & controls. Examples of PAS staining & immunostaining with gamma H2AX (green) & WT1 (red), pATM & 5mC in glomeruli of IgA nephropathy & controls. (A) A control kidney sample of 44-year-old female, (B) 65-year-old male of IgA nephropathy without podocytopathic features & (C) 55-year-old male of IgA nephropathy with podocytopathic features. Arrows indicate gamma H2AX & WT1 double-positive cells. Scale bars: 50 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31937846), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Histone H2AX [p Ser139] Antibody [NB100-384] - UHRF1 deficiency resulted in impaired meiotic recombination & defective pachynema.a Double immunofluorescence of SYCP3 (green) & DMC1 (red) in testicular spread preparations. b, c The number of DMC1 foci in zygotene stage (b) & pachytene stage (c). d Immunostaining for SYCP3 (red) & gamma H2AX (green). e The percentage of abnormal gamma H2AX foci in the pachytene stage. f Immunostaining for SYCP3 (red) & MLH1 (green). g The number of MLH1 foci in pachynema. h Immunostaining for SYCP3 (red) & H1t (green). i The percentage of spermatocytes with H1T staining. ***p ≤ 0.001; *p ≤ 0.05. Scale bar, 5 μm in a, d, f, h. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32081844), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Histone H2AX [p Ser139] Antibody [NB100-384] - mTORC1/2 activity prevents Cisplatin-induced cell death in MCF-10A cells. (A) Western blot displaying effects on mTOR signaling during a dose escalation of PP242 treatment in MCF-10A cells; (B) Western blot displaying effects of mTOR signaling on a dose escalation of cisplatin treatment in MCF-10A cells; (C) Western blot displaying effects on mTOR signaling & cell death during non-treated, Cisplatin, PP242, & Cisplatin + PP242-treated MCF-10A cells. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31771139), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Histone H2AX [p Ser139] Antibody [NB100-384] - Pharmacologic inhibition of ATM rescues oxidative stress-induced senescence by suppressing ATM- & NEMO-mediated NF-kappa B activation. (A) Representative images of primary WT & Ercc1-/- MEFs were induced to undergo senescence by serial passaging at 20% oxygen. At passage 5, MEFs were grown in the presence or absence of KU-55933 (10 μM) for 72 hrs. Senescence was determined by SA-beta gal staining. Images were obtained at the magnification of 10x. (B) Quantitation of the percent SA-beta gal positive cells. Graph represents the mean +/- s.e.m. of three independent experiments. Student’s t-test, ***p <0.001, ****p <0.0001. (C) Passage 5 Ercc1-/- MEFs treated with vehicle or KU-55933 (10 μM) for 72 hours were collected & levels of p21 & p16INK4a were determined by western blotting. (D) Passage 5 Ercc1-/- MEFs were treated with KU-55933 (10 μM) for 72 hours & whole cell lysate (CL) & nuclear extracts (NE) were analyzed by immunoblotting for expression of proteins involved in the DNA damage response. (E) Whole cell lysate (CL) & nuclear extract (NE) were extracted from Ercc1-/- MEFs treated with 10 μM of KU-55933 for analysis of nuclear NEMO & p65. GAPDH was used as a loading control of total proteins & LaminA/C as a loading control of nuclear protein. (F) Passage 5 WT & Ercc1-/- MEFs transfected with a NF-kappa B-luciferase reporter construct were cultured in the presence or absence of KU-55933 (10 μM) & were collected for luciferase assays after 72 hours. (G) qRT-PCR analysis of mRNA expression in passage 5 WT & Ercc1-/- MEFs treated with or without of KU-55933 (10 μM) for 72 hrs. P values were determined using a Student’s t-test. *p<0.05, **p<0.01, ***p <0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32201398), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: Histone H2AX [p Ser139] Antibody [NB100-384] - Orally administered lemongrass & white tea extract reduce tumor size in lymphoma xenograft model in immunocompromised miceImmunocompromised mice were subcutaneously injected with cancerous cells & tumors were allowed to establish. Treatments occurred every other day & the studied compound or the equivalent vehicle control administered orally for three weeks. (A, B) The tumors were photographed before & after extraction from the animals. (C) Tumor volume & mass were measured two times per week. (D) Immunohistochemistry analysis of sectioned tumor tissues from the lymphoma study. Each section was subjected to the specified antibody followed by a biotinylated secondary antibody. Detection was done using a DAB Peroxidase HRP Substrate Kit (brown) followed by Hematoxylin counterstaining (purple). Images were obtained using inverted bright field microscopy. Sectioning results are representative of three individual tumors. Scale bar is 50 microns. Statistical analysis using One-Way ANOVA. *p < 0.05 vs tumor volume of the control. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.22502), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Histone H2AX [p Ser139] Antibody [NB100-384] - CPT induces transcription- & proteasome-dependent apoptosis in quiescent WI38 hTERT cells. (A & B) Serum-starved cells were treated with DMSO or FLV (1 μM) for 1 h before the addition of DMSO or CPT (25 μM) for 16 & 24 h. (A) Percentages of cells that remained attached to culture flasks (means ± SD of quadruplicates). (B) Cell survival was analyzed by a CellTiter-Blue assay (means ± SD of triplicates). (C–G) Western blot of the indicated proteins in serum-starved cells treated for 1 h with DMSO or with FLV (1 μM) (C), lactacystin (10 μM) (D), veliparib (5 μM) or olaparib (10 μM) (E), ATMi (10 μM) (F) or DNA-PKi (10 μM) (G) before the addition of DMSO (‘-’, 24 h in panels C & F) or CPT (25 μM) for the indicated times. Data shown are representatives from two to three experiments. PARPCL: cleaved PARP. H2AX & alpha Tubulin were the loading controls. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26578593), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Histone H2AX [p Ser139] Antibody [NB100-384] - Inhibition of DNA-PK prevents monoubiquitination of H2AX & H2A in CPT-treated quiescent WI38 hTERT cells. (A–E) Serum-starved cells were treated with DMSO or DNA-PKi (10 μM) for 1 h before the addition of DMSO (untreated) or CPT (25 μM) for 1 h. (A) Western blot of gamma H2AX. +Ub1 indicates gamma H2AX monoubiquitinated. The top panel shows quantification of Ub1-gamma H2AX normalized to alpha Tubulin (means ± SEM, n = 4). **P < 0.01. (B & C) Cells were pre-extracted with CSK buffer before co-staining for Ub-H2A (red) & 53BP1 phosphorylated on S1778 (p53BP1) (green). (B) Representative pictures. Images were merged to determine colocalization (yellow). The large Ub-H2AX focus at the periphery of the nuclei of untreated & CPT-treated cells may marks the inactive X chromosome as reported (91). (C) Percentages of nuclei with at least 5 Ub-H2A foci (means ± SEM, n = 3, 100 nuclei were analyzed for each treatment in each experiment). ***P < 0.001. (D & E) Cells were co-stained for ubiquitinated proteins (FK2, red) & gamma H2AX (green). (D) Representative pictures. Images were merged to determine colocalization (yellow). (E) Number of FK2 foci per nucleus from one representative experiment (76–111 nuclei were analyzed for each treatment) out of three. ****P < 0.0001. In the microscopic images, nuclear contours, identified by DAPI staining (blue in the merge images at bottom), are indicated by dashed lines. Bars: 10 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26578593), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Histone H2AX [p Ser139] Antibody [NB100-384] - The production of DSBs depends on Top1 degradation in CPT-treated quiescent cells. (A–C) Serum-starved WI38 hTERT cells were co-transfected with siRNAs against cullin 3 & cullin 4B or against a control sequence & then treated with DMSO (−CPT) or 25 μM CPT (+CPT) for 1 h. (A & B) Western blotting of the indicated proteins. alpha Tubulin: loading control. (C) Number of gamma H2AX foci per nucleus from one representative experiment (246–348 nuclei were analyzed for each treatment) out of three. ***P < 0.001. (D & E) Serum-starved WI38 hTERT cells were treated with DMSO or MG132 (50 μM) for 1 h before exposure to 0.8 Gy IR. One hour post-irradiation, cells were co-stained for gamma H2AX (green) & 53BP1 (red). (D) Representative pictures. (E) Number of gamma H2AX foci per nucleus from one representative experiment (162–180 nuclei were analyzed for each treatment) out of three. Ns: not significant. (F & G) U2OS EV28 cells were treated with DMSO or MG132 (10 μM) for 1 h before the addition of ethanol (untreated) or 300 nM 4-hydroxitamoxifen (4OHT) for 4 h to express AsiSI in the nucleus (42). (F) Representative pictures of cells co-stained for gamma H2AX (green) & 53BP1 (red). (G) ChIP analysis using an anti-gamma H2AX antibody (black) or a non-immune antibody (IgG, gray). Enrichment was assessed by QPCR amplification using primers proximal to two AsiSI sites located inside two genes (Gene I: SFRS6, Gene II: CCD47) & primers distal to an AsiSI site (Control). Enrichment was normalized to the maximum recovery for each experiment (means ± SEM, n = 3). Ns: not significant; *P < 0.05. In the microscopic images, nuclear contours, identified by DAPI staining (not shown), are indicated by dashed lines. Bars: 10 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26578593), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Histone H2AX [p Ser139] Antibody [NB100-384] - Inhibition of DNA-PK prevents monoubiquitination of H2AX & H2A in CPT-treated quiescent WI38 hTERT cells. (A–E) Serum-starved cells were treated with DMSO or DNA-PKi (10 μM) for 1 h before the addition of DMSO (untreated) or CPT (25 μM) for 1 h. (A) Western blot of gamma H2AX. +Ub1 indicates gamma H2AX monoubiquitinated. The top panel shows quantification of Ub1-gamma H2AX normalized to alpha Tubulin (means ± SEM, n = 4). **P < 0.01. (B & C) Cells were pre-extracted with CSK buffer before co-staining for Ub-H2A (red) & 53BP1 phosphorylated on S1778 (p53BP1) (green). (B) Representative pictures. Images were merged to determine colocalization (yellow). The large Ub-H2AX focus at the periphery of the nuclei of untreated & CPT-treated cells may marks the inactive X chromosome as reported (91). (C) Percentages of nuclei with at least 5 Ub-H2A foci (means ± SEM, n = 3, 100 nuclei were analyzed for each treatment in each experiment). ***P < 0.001. (D & E) Cells were co-stained for ubiquitinated proteins (FK2, red) & gamma H2AX (green). (D) Representative pictures. Images were merged to determine colocalization (yellow). (E) Number of FK2 foci per nucleus from one representative experiment (76–111 nuclei were analyzed for each treatment) out of three. ****P < 0.0001. In the microscopic images, nuclear contours, identified by DAPI staining (blue in the merge images at bottom), are indicated by dashed lines. Bars: 10 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26578593), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Histone H2AX [p Ser139] Antibody [NB100-384] - DDR & NF-kappa B are activated concomitantly in senescent MEFs & aged tissues. (A) Immunoblot detection of p-p65 & total p65 in liver tissue from 16-week-old WT (n=3) & Ercc1-/ delta (n=3) mice. (B) Immunoblot detection of phosphorylation of ATM & downstream targets gamma H2AX & p21 in liver from 16-week-old WT & Ercc1-/ delta mice. (C) Immunoblot detection of phosphorylation of NF-kappa B & I kappa B alpha in liver lysates from 3, 12 & 24 month-old WT mice. n=3 mice per group. (D) Immunoblot detection of p-ATM, ATM & p21 in the same liver lysates. (E) Immunoblot detection of DDR effectors in nuclear extracts from passage 5 WT & Ercc1-/- MEFs, grown at 20% oxygen. (F) Level of NF-kappa B activation is higher in Ercc1-/- MEFs compared to WT MEFs at passage 5, as measured by Immunoblot detection of p-p65 & total p65 in WT & Ercc1-/- MEFs at passage 5 after culturing in 20% oxygen. (G) Representative images of immunofluorescent detection of p65 & NEMO in passage 4 WT & Ercc1-/- MEFs grown at 20% oxygen. Blue: DAPI staining; Green: p65 (top panel) or NEMO (bottom panel). Images were taken at the magnification of 60x. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32201398), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Histone H2AX [p Ser139] Antibody [NB100-384] - Induction of ubiquitin/proteasome-dependent DSBs in CPT-treated quiescent WI38 hTERT cells. (A–F) Serum-starved cells were treated with DMSO (1 h) or with MG132 (50 μM, 1 h), lactacystin (10 μM, 1 h), bortezomib (1 μM, 4 h), G5 (1.5 μM, 0.5 h) or Pyr-41 (9 μM, 0.5 h) before the addition of DMSO (untreated) or 25 μM CPT for 1 h & then co-stained for gamma H2AX (green) & 53BP1 (red) or analyzed by Western blot. ‘-’ in panels C & F means cells treated with DMSO. (A & D) Representative pictures. Nuclear contours, identified by DAPI staining (not shown), are indicated by dashed lines. Bars: 10 μm. (B & E) Number of gamma H2AX foci per nucleus from two independent experiments (147–153 nuclei were analyzed for each treatment). ****P < 0.0001. (C & F) Western blot of gamma H2AX. alpha Tubulin: loading control. Dashed lines indicate that intervening wells have been spliced out. The top panels show quantification of gamma H2AX normalized to alpha Tubulin (means ± SEM, n = 4 in panel C, n = 3 in panel F). ***P < 0.001; **P < 0.01. (G & H) Detection of DSBs by neutral Comet assays in serum-starved cells treated with DMSO or MG132 (25 μM) for 1 h before the addition of DMSO (untreated) or CPT for 1 h (7.5 μM for experiments (Exp) I & II; 5 & 7.5 μM for Exp III). (G) Representative pictures of nuclei from Exp I. (H) Quantification of neutral Comet tail moments for three independent experiments (95–133 nuclei were analyzed for each treatment in each experiment). ***P < 0.001; ****P < 0.0001. The untreated & CPT data from Exp I are from the same experiment as that of Supplementary Figure S3D. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26578593), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Histone H2AX [p Ser139] Antibody [NB100-384] - Curcumin Analogs Induce Apoptosis in Cancerous Cells by Several Pathways. (a) E6-1 (Jurkat), dominant negative FADD (dnFADD) Jurkat, & overexpressing BCL-2 Jurkat were treated for 48 hours then stained for Annexin V & PI (b) E6-1 cells were plated & treated with or without the broad spectrum caspase inhibitor ZVAD(oMe)-FMK for 48 hours. Cells were stained for Annexin V & PI. Results were obtained using image-based cytometry with the Y-axis representative of percent of cells positive for Annexin V (green), PI (red), Annexin V & PI (yellow), or negative for both Annexin V & PI (blue). Values are expressed as a mean ± SD from three independent experiments. (c) E6-1 cells were treated for 24 hours with or without the broad spectrum caspase inhibitor ZVAD(oMe)-FMK & the studied compounds, lysed & subjected to Western blot analysis. (d) NHF & NCM460 cells were treated for 48 hours & 72 hours respectively, lysed & subjected to Western blot analysis. Bands were visualized with a chemiluminescence reagent. Images are representative of three independent experiments. Statistical calculations were performed using Two-Way ANOVA multiple comparison. *p < 0.05 vs % viable of Control (DMSO); #p < 0.05 vs % viable cells for groups without Z-VAD(oMe)FMK. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28439094), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Ca

Species Glossary

Reactivity: Hu, Mu, Rt, Ca

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-384AF350
Alexa Fluor 405	NB100-384AF405
Alexa Fluor 488	NB100-384AF488
Alexa Fluor 532	NB100-384AF532
Alexa Fluor 594	NB100-384AF594
Alexa Fluor 647	NB100-384AF647
Alexa Fluor 700	NB100-384AF700
Alexa Fluor 750	NB100-384AF750
Allophycocyanin	NB100-384APC
Biotin	NB100-384B
CoraFluor 1	NB100-384CL1
DyLight 350	NB100-384UV
DyLight 405	NB100-384V
DyLight 488	NB100-384G
DyLight 550	NB100-384R
DyLight 594	NB100-384DL594
DyLight 650	NB100-384C
DyLight 680	NB100-384FR
DyLight 755	NB100-384IR
FITC	NB100-384F
HRP	NB100-384H
Janelia Fluor 525	NB100-384JF525
Janelia Fluor 549	NB100-384JF549
Janelia Fluor 646	NB100-384JF646
Janelia Fluor 669	NB100-384JF669
PE	NB100-384PE
PerCP	NB100-384PCP
mFluor Violet 450 SE	NB100-384MFV450
mFluor Violet 500 SE	NB100-384MFV500
mFluor Violet 610 SE	NB100-384MFV610

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NB100-2284

CD31/PECAM-1 Antibody

(5 Reviews)

(151 Publications)

0.1 ml

NB100-2284

Staining of CD31/PECAM-1 in Rat Liver sample. IHC-P image submitted by a verified customer review.

Human lung adenocarcinoma using. Antibody at 1:100 with citrate epitope retrieval at pH 6.0.

Analysis in rat muscle blood vessels using. IHC-P image submitted by a verifed customer review.

Matrix-HA supplemented with strontium was implanted subcutaneously in mice, where immunochemistry analysis and histology occurred of the newly formed tissues. (C) Within each matrix: Matrix-HA, Matrix-8Sr-HA and Matrix-50Sr-HA, were indicated by immunostaining of CD31 for the newly formed tissues. (D) Number of vessels within each tissue was quantified using NDP view software. Unit is square mm. Immunostaining analysis of slides occurred for 2 samples per condition and 3 sections were analyzed per sample and per group of matrix. Citation: Ehret C, Aid-Launais R, Sagardoy T, Siadous R, Bareille R, Rey S, et al. (2017) Strontium-doped hydroxyapatite polysaccharide materials effect on ectopic bone formation. PLoS ONE 12(9): e0184663. //doi.org/10.1371/journal.pone.0184663

Imaging of HUVEC monolayer on glass substrate. Fixation with 4% PFA. Blocking with Goat Serum at 1:100. Incubation 2 hours at room temperature. ICC/IF image submitted by a verified customer review.

Analysis of canine kidney tissue using CD31/PECAM-1 antibody. Antibody was used at a 1:50 concentration in Casein in PBS and left at 4C overnight on paraffin embedded canine bladder tissue. HIER was performed in Tris/EDTA buffer, pH 9 for two hours at 75C. Image from verified customer review.

Analysis in pig SC cells using CD31/PECAM-1 Antibody (red). Blue color showing nucleus labeling. ICC/IF image submitted by a verified customer review.

Immunohistochemistry evaluation of CD31/PECAM-1 expression. Representative images of CD31/PECAM-1 staining of vessels in the three groups of implanted matrices (scale bar = 100 um). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-06258-0) licensed under a CC-BY license.

CD31 staining on E11.5 mouse Pharyngeal Mesoderm. Fixed with 4% PFA overnight. Blocked with 1% BSA. Primary antibody at 1:100. Secondary antibody at 1:1000, conjugated to Alexa Fluor 488. IHC-Fr image submitted by a verfied customer review.

Immunohistochemistry evaluation of CD31 expression. (a) Representative images of CD31 staining of vessels in the three groups of implanted matrices (scale bar = 100 µm). (b) Quantification of vessel density inside the implanted matrices at 5 weeks post implantation (n = 6; Average ± SD). NS and **denote Non Significant and p < 0.01, respectively.

Immunohistochemistry: Rabbit Polyclonal CD31/PECAM-1 Antibody [NB100-2284] - Staining of CD31 in human lung tissue. Nuclei in blue and CD31 (vessel) in green. Mouse anti-CD31 (1:100, overnight incubation at +4°C), followed by donkey anti-mouse 488 (1:400, 3h at RT). Image from a verified customer review.

Immunohistochemistry: CD31/PECAM-1 Antibody [NB100-2284] - Antibodies selected for characterization of stromal cells & vasculature in FFPE specimens are species-specific. (A, B) Labeling of a patient colon tumor (A) & Colo205 human cell line xenograft (B) with anti-human mitochondrial antibody (single arrow- tumor cells; double arrows- stroma). (C, D) Labeling of a patient pancreatic tumor (C) & Colo205 human cell line xenograft (D) with anti-human CD34. (E, F) Labeling of a patient lung tumor (E) & Colo205 xenograft (F) with anti-human CD31. (G, H) Labeling of a patient lung tumor (G) & Colo205 xenograft (H) with anti-mouse CD34. (bars = 50 μ). Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: CD31/PECAM-1 Antibody [NB100-2284] - At the time of second passage, vessels in successfully growing xenografts of four different tumor types surveyed were of murine origin: A-C Colon; D-F Lung; G-I Pancreatic; J-L Renal Cell Carcinoma. For each tumor type, a representative section of an original patient specimen labeled with anti-huCD31 is shown (A, D, G, J). For each tumor, sections of the first passage xenografted tumor when it was resected are also shown; while no huCD31(+) vessels were identified in the xenografts (B, E, H, K), msCD34(+) vessels were abundant (C, F, I, L). (bars = 50 μ). Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: CD31/PECAM-1 Antibody [NB100-2284] - Vascularization of an engrafted patient colon tumor. A patient colon tumor was implanted in a cohort of mice & vessel development was analyzed over 8 weeks. Vessels in the original patient specimen labeled strongly for huCD31 (A) & not for msCD34 (D). Representative sections showing loss of huCD31(+) vessels (B- 4 weeks, C- 7 weeks) & presence of msCD34(+) vessels (E- 4 weeks, F- 7 weeks) are shown. The graph (G) summarizes this process; huCD31(+) vessels were rapidly lost, & by one week, mouse vessels were the predominate vessels present in the colon tumors (no data for msCD34 at 2 weeks; bars = 100 μ). Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: CD31/PECAM-1 Antibody [NB100-2284] - KCl pulse decreased membrane, but increased nuclear, detection of NHE1 in b.End3 endothelial cells.(A) Representative immunofluorescence images of bEnd.3 cells at two time-points after KCl or aCSF pulse. (B) Representative immunoblots of NHE1, alpha -tubulin, PECAM, & lamin B in cytosol, membrane, & nuclear fractions of bEnd.3 cells harvested at 5 min after KCl or aCSF pulse. (C = cytosol, M = membrane, N = nuclear) Values represent the mean ratio of NHE1 detection ± SEM (n = 11–12). (C) Representative immunoblots indicating NHE1 & alpha -tubulin as a loading control in whole cell lysate of bEnd.3 cells harvested at 5 min after KCl or aCSF pulse. Values represent the % of aCSF-treated relative expression ± SEM (n = 6). (D) Representative immunoblots of NHE1 & alpha -tubulin as a loading control in whole lysate of microvessels harvested at 90 min after cortical injection of KCl or aCSF. Values represent the % of naive relative expression ± SEM (n = 6). # denotes significantly different vs naïve (p<0.01), as assessed by one-way ANOVA (E) Intracellular pH during aCSF or KCl pulse. All data represent mean ± SEM (n = 45). (F) Extracellular pH during aCSF or KCl pulse. All data represent mean ± SEM (n = 6) in triplicate. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32469979), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Immunocytochemistry/ Immunofluorescence: CD31/PECAM-1 Antibody [NB100-2284] - Impact of T‐AuNPs on endothelial cells in vivo. Immunofluorescent staining for endothelial cells using an anti‐CD 31 antibody revealed an intact endothelial cell monolayer in the uninjured control carotid artery (N = 3). As expected post balloon angioplasty, there is no endothelial cell monolayer of the injured left carotid arteries in the injury alone, injury + T‐AuNP, & injury + S‐AuNP treatment conditions. Green indicates autofluorescence of the elastic lamina. Magenta indicates endothelial cells. Images obtained using 25× magnification with exposure time of 400 msec. TUNEL staining for apoptosis revealed no evidence of endothelial cell apoptosis in the uninjured control carotid arteries, with a prominent endothelial cell monolayer (white arrowhead). Mild apoptosis was noted in the media of balloon‐injured left carotid arteries, as expected. Black arrows indicate apoptotic cells. Representative images obtained using 40× magnification. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28242820), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: CD31/PECAM-1 Antibody [NB100-2284] - The growth of a patient mesothelioma xenograft was supported by development of a murine vascular network. A patient mesothelioma was implanted in a cohort of mice & monitored for tumor growth. Once tumors began to actively grow, representative tumors were resected at weekly intervals & analyzed for vessel content. Staining for huCD31 was prominent in the original patient specimen (A), much reduced at 4 weeks (B) & negligible at 9 weeks (C). In contrast, patient specimens were not stained for msCD34 (D), whereas at 4 weeks large numbers of vessels stained for msCD34 (E) & by 9 weeks, msCD34 labeled vessels were predominant (F). The graph (G) summarizes the loss of detectable human vessels & acquisition of murine vessels over a 9 week period. (bars = 100 μ). Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: CD31/PECAM-1 Antibody [NB100-2284] - TGF‐ beta 1 released by MSCs contributes to induced myofibroblast differentiation & granulation tissue formationA–C2.5 × 105 of TGF‐ beta 1 siRNA or control siRNA‐transfected AT‐MSCs were intradermally injected around each of CD18−/− murine wound. PBS mock injection served as negative control. Wound tissue was harvested at day 2, 5, & 7 post‐wounding for quantification of human TGF‐ beta 1 mRNA (A) at day 2 by qPCR, total TGF‐ beta 1 (B), & active TGF‐ beta 1 (C) protein at day 5 by ELISA. Data are expressed as mean ± SEM, n = 3 wounds per group, **P < 0.01, by one‐way ANOVA with Tukey's test.D–GExpression of alpha ‐SMA (D & E) & CD31 (F & G) at days 5 & 7 by immunostaining on tissue sections. The dashed lines indicate the border of the wound bed & epidermis or eschar. e, epidermis; es, eschar; wb, wound bed. Scale bars: 100 μm. Quantification data are expressed as mean ± SEM, n = 3 wounds per group, *P < 0.05, ***P < 0.001, by one‐way ANOVA with Tukey's test.Source data are available online for this figure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32080965), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: Rabbit Polyclonal CD31/PECAM-1 Antibody [NB100-2284] - CD31 positive vessels in a canine splenic hemangiosarcoma.Tissue slides were loaded into automated research stainer, dewaxed and pretreated with EDTA-based epitope retrieval ER2 solution for 20 mins at 100°C. After a peroxide incubation for 5 min, the rabbit antibody against CD31 (NB100-2284) (1:100) was incubated for 60 mins at room temperature followed by Bond Polymer (anti-rabbit HRP) incubation for 8 min at room temperature. Mixed DAB reagent (Polymer Refine Detection Kit) was incubated for 10 mins, and Hematoxylin (Refine Detection Kit) counterstaining for 10 mins. After staining, sample slides were washed in water, dehydrated using ethanol gradient (70%, 90%, 100%), washed three times, and mounted in mounting medium. Image from a verified customer review.

Reactivity:

Hu, Mu, Rt, Po, Ca

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ca

Applications:

, ,

Applications: , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

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NB300-213

MAP2 Antibody - BSA Free

(5 Reviews)

(132 Publications)

0.05 ml

NB300-213

Mouse primary cultures stained with MAP2 antibody. ICC/IF image submitted by a verified customer review.

Immunofluorescent analysis of cortical neuron-glial cell culture from E20 rat stained with chicken pAb to microtubule associated protein 2 (MAP2), NB300-213, dilution 1:10,000 in red, and mouse mAb to MAP-tau, NPB2-25162, dilution 1:2,000, in green. The blue is DAPI staining of nuclear DNA. NB300-213 antibody stains dendrites and perikarya of neurons, while NBP2-25162 antibody labels neuronal perikarya, dendrites and also axonal process. As a result perikarya and dendrites appears orange-yellow, since they contain both MAP2 and tau, while axons are green.

Western blot analysis of whole brain tissue lysates using chicken pAb to microtubule associated protein 2 (MAP2), NB300-213, dilution 1:50,000 in green: [1] protein standard (red), [2] adult rat brain, [3] embryonic E20 rat brain, [4] adult mouse brain. Strong band observed at ~280 kDa mark corresponds to two major isoforms of MAP2 protein referred to as MAP2A and MAP2B. Smaller fragments of these isoforms are also detected if the antibody is used at higher concentrations.

PEA decreases astrocyte activation in organotypic cultures of rat hippocampi and rescues neuronal CA3 damage caused by Abeta challenge. Abeta-challenged (1 ug/ml) slices of rat hippocampi were treated for 24 hours with PEA (0.1 uM) in the presence of the selective PPAR? antagonist (GW9662, 9 nM) or the selective PPARalpha antagonist (MK886, 3 uM). Representative photomicrographs of the CA3 region showing the results of immunofluorescence experiments aimed at investigating the effect of treatments on astrocyte activation and neuronal loss, as determined by immunostaining for GFAP (green) and MAP2 (red) alone or merged, respectively. Nuclei were stained with Hoechst (blue). Scale bar: 10 um. Arrows in the photomicrographs indicate astrocyte infiltration events and apoptotic condensation in the nuclei of adjacent neurons.

Human iPS derived neurons labeled with MAP2 (in green) at 1:5000 dilution. Image from verified customer review.

MAP2 in the retina detected by NB300-213 antibody (magenta). DAPI (blue). Image from verified customer review.

Human DA neurons 28 DIV stained with MAP2 antibody (blue). ICC/IF image submitted by a verified customer review.

PEA inhibits astroglial proliferation and reduces neuronal loss in mixed neuroglia co-cultures exposed to Abeta. Abeta-challenged (1 ug/ml) astrocyte/neuron mixed cultures were treated with PEA (0.1 uM) i9662, 9 nM) on the presence of the selective PPAR-gamma antagonist (GWr the selective PPARalpha antagonist (MK886, 3 uM). After 24 hours of treatment, cells were processed for analyses. Immunofluorescence photomicrographs showing the effect of the treatments on astrocyte proliferation and neuronal loss, as determined by immunostaining for GFAP (green) and MAP2 (red), respectively. Arrows indicate chromatin condensation in nuclei stained with Hoechst dye (blue) as markers of apoptotic events. Scale bar: 10 um. Image collected and cropped by CiteAb from the following publication (//jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094-9-49), licensed under a CC-BY license.

Oxytocin receptors (Oxtr) are expressed in mouse hippocampus. Confocal images of Oxtr-expressing neurons in the CA1 subfields of the medial and caudal hippocampus from OxtrVn/+ sections, triple stained with antibodies directed against NeuN (red), MAP2 (magenta), and Venus (green). Image collected and cropped by CiteAb from the following publication (//elifesciences.org/articles/22466), licensed under a CC-BY license.

Differential and neuron-specific Oxtr expression in mouse hippocampus. Immunohistochemical mapping of Venus expression in coronal sections of the rostral, medial, and caudal hippocampus from three weeks old OxtrVn/+ mice triple-labeled with antibodies against NeuN (red), Venus (green), and Map2 (magenta). Image collected and cropped by CiteAb from the following publication (//elifesciences.org/articles/22466), licensed under a CC-BY license.

ATM activation by chloroquine alleviates senescence.(a) Immunoblots showing protein levels of ATM, NBS1, and RAP80 in human skin fibroblasts (HSFs). A gradually increased level of p16 indicates cellular senescence, while elevated gamma H2AX level indicates accumulated DNA damage. (b) Immunoblots showing protein levels of ATM, NBS1, and RAP80 in mouse embryonic fibroblasts (MEFs). (c) Immunoblots showing protein levels of ATM, NBS1, and RAP80 in brain tissues isolated from 3-, 10-, and 18-month-old male mice. (d) SA-beta -Gal staining in HSFs treated with sh-ATM or scramble shRNA. Scale bar, 100 µm. (e) Quantification of SA-beta -Gal-positive staining of (d) from five views randomly captured for each group. Data represent means ± SEM. ***p<0.001. (f) Immunoblots showing increased gamma H2AX and unaffected LC3I/II in HSFs treated with sh-ATM or scramble shRNA. (g) Immunoblots showing protein levels of pS1981 ATM, gamma H2AX, and cleaved caspase-3 in HSFs treated with 10 μM of CQ for indicated time. (h) SA-beta -Gal staining in HSFs expressing either scramble or ATM shRNA treated with 1 μM CQ or DMSO (12 hr). Scale bar, 100 µm. (i) Quantification of SA-beta -Gal-positive staining of (h) from five views randomly captured for each group. Data represent means ± SEM. ***p<0.001; ‘N.S.’ indicates no significant difference. (j) HSFs at passage 20 were continuously cultured with 1 μM CQ or DMSO, and cell number was calculated at each passage. Data represent means ± SEM. ***p<0.01. (k) Immunoblots showing protein levels of gamma H2AX, p62, and LC3 in MEFs treated with 1 μM CQ or DMSO. Note that CQ had little effect on the expression levels of p62 and LC3. (l) MEFs at passage one were continuously cultured in 20% O2 with 1 μM CQ or DMSO, and cell number was determined at each passage. Data represent means ± SEM. ***p<0.01.10.7554/eLife.34836.006Figure 1—source data 1.Statistical analysis for SA-beta -Gal positive staining.10.7554/eLife.34836.007Figure 1—source data 2.Statistical analysis for EdU positive staining.Statistical analysis for SA-beta -Gal positive staining.Statistical analysis for EdU positive staining.Decline of ATM-centered DNA repair machinery during senescence.(a) Real-time PCR analysis showing progressively elevated mRNA level of p21 in continuously cultured human endothelial cells (HUVEC). **p<0.01. (b) SA-beta -Gal staining of HUVEC cells at indicated passages. Scale bar, 100 µm. (c) HUVEC cells at P21, P18, P12, and P7 were subjected to transcriptome analysis. A minimum average rpkm value of 1.0 and maximum 10% fluctuation in young cells (P7 Vs P12) was set as the threshold. Genes were downregulated by more than 20% in pre-senescent, and senescent cells compared with young cells (P21/P18 Vs P12/P7) were selected. (d) Pathway analysis of genes identified in (c) by STRING v10. (e) Downregulation of ATM-related DNA repair genes during senescence.ATM regulates replicative senescence.(a) Representative images showing cells treated with Scramble (sh-NC) or sh-ATM. (b) Percent EdU-positive cells in sh-NC or sh-ATM treated HSFs. Views were randomly captured and at least 100 cells were included in each group. Data represent means ± SEM. ***p<0.001. (c) Immunoblots showing protein levels of pS1981 ATM and gamma H2AX in HSFs treated with 10 μM chloroquine (CQ) or 0.4 μM CPT (4 hr). Note that CQ activated ATM (pS1981) without increasing gamma H2AX, while CPT activated ATM accompanied by increased gamma H2AX. (d) SA-beta -Gal staining in primary MEFs treated with 1 μM CQ or DMSO. Scale bar, 100 µm. (e) Quantification of SA-beta -Gal-positive staining of (d) from five views randomly captured for each group. Data represent means ± SEM. ***p<0.001. (f) Percent EdU-positive cells in HSFs treated with DMSO, 1 μM or 10 μM CQ. Views were randomly captured and at least 100 cells were included in each group. Data represent means ± SEM. ***p<0.001. (g) Representative images showing proliferative HSFs treated with different doses of CQ for the indicated time points. (h) Immunoblots showing LC3B levels in HSFs treated with indicated dose of CQ for indicated period of time. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29717979), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: MAP2 Antibody [NB300-213] - SUMO1 conjugates are not localized to amyloid plaques. (a) Sagittal brain sections of 24‐week‐old KI/AD & WT/AD animals were stained using anti‐HA antibodies (red), 6E10 antibodies (green) that label amyloid beta 1–42 among other amyloid beta variants (epitope lies within amino acids 3–8 of amyloid beta) & anti‐MAP2 antibodies (blue). Sections of the hippocampal subiculum are shown. Images are representatives of three independent experiments. Scale bar, 25 μm. (b) Anti‐HA signal intensity in amyloid plaques & in cell nuclei was quantified using ImageJ (N = 3, ***significance between WT/AD & KI/AD, p = .0007 in Student's t test) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29633471), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: MAP2 Antibody [NB300-213] - Oxytocin receptors (Oxtr) are expressed in mouse hippocampus.(a & b) Coronal sections of the hippocampus (a) & striatum (b) from OxtrVn/+ & Oxtr+/+ mice stained for Venus & DAPI (ac, anterior commisure; cx, cortex; hip, hippocampus; str, striatum). (c) Confocal images of rostral Oxtr+/+ & OxtrVn/+ brain sections including hippocampus, double-labeled for Venus (green) & NeuN (red). (d) Confocal images of Oxtr-expressing neurons in the CA1 subfields of the medial & caudal hippocampus from OxtrVn/+ sections, triple stained with antibodies directed against NeuN (red), MAP2 (magenta), & Venus (green). (e) Representative images of hippocampal mass cultures obtained from OxtrVn/+ mice at P0 mice & processed for immunofluorescence staining at 5, 10, & 14 DIV using antibodies to Venus (green) & GAD67 (red), & DAPI (blue). (f) Quantitative analysis of Venus-positive neurons with (GAD67+) & without colabeling for GAD67 (GAD67-) in hippocampal mass cultures obtained from OxtrVn/+ mice at P0 mice & processed for immunofluorescence staining at 5, 10, & 14 DIV. Data are shown as mean ± SEM.DOI://dx.doi.org/10.7554/eLife.22466.015 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28231043), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Immunocytochemistry/ Immunofluorescence: MAP2 Antibody [NB300-213] - SUMO1 conjugates remain nuclear during increased amyloid burden. (a) Western blot analysis of subcellular fractions of 36‐week‐old KI/WT & KI/AD mouse brain using anti‐HA antibody (upper two panels) & antibodies to GluN1 (a marker of the postsynaptic compartment) & Synaptophysin (a marker of the presynaptic compartment) to validate the fractionation procedure (lower two panels). H, homogenate; P1, nuclear pellet; S1, supernatant after P1 sedimentation; P2, crude synaptosomal pellet; S2, supernatant after P2 sedimentation; LP1, lysed synaptosomal membranes; LS1, supernatant after LP1 sedimentation; LP2, pellet after LS1 sedimentation, SPM, synaptic plasma membranes. Bracket indicates the anti‐HA signal representing SUMOylation, arrow indicates RanGAP1, stars indicate nonspecific signal detected by the anti‐HA antibody. (b) Brain sagittal sections of aged (36 weeks old) KI/AD (left panels) & WT/AD (right panels) mice were immunostained using antibodies directed against HA (red, labels HA‐HA conjugates), MAP2 (green, labels neuronal somata & dendrites), & Synapsin1 (Syn1, magenta, labels synapses). The images show triple‐labeled neurons of hippocampal subiculum (b), cortical layer 5 (c), & proximal apical dendrite from hippocampal CA3 (d). The white line shows the orientation of the line‐scan used to generate the image stack shown in the bottom side view. Scale bar: 10 μm. Note that the anti‐HA immunosignal is mainly located in neuronal nuclei, only background staining is observed in WT/AD mice. Little anti‐HA signal is observed along MAP2‐positive structures & does not colocalize with Synapsin1 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29633471), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Immunocytochemistry/ Immunofluorescence: MAP2 Antibody [NB300-213] - Validation of the CRISPR/Cas9 Alsin-/- cells in iPSC, NPC & iPSC-sMN.(A) Electrophoresis result of the PCR reaction using primers flanking exon three & within exon 3. Homozygous deletion was confirmed by the absence of a 2.8 kb. (B) Protein lysates from WT KOLF & Alsin-/- iPSC were loaded onto SDS-PAGE & immunoblotted with Alsin antibody. A band detected at ~184 kDa, but absent in Alsin-/-, corresponds to full-length Alsin. (C) WT KOLF, Alsin-/- iPSC, & neuroprogenitor cells (NPC) were immunostained with pluripotency markers Oct4 & Lin28, & neuroprogenitor markers Sox2 & Pax6, respectively. (D–E) WT KOLF & Alsin-/- iPSC-sMN were immunostained with motor neuron markers such as ChAT, HB9, & ISL1, nuclear dye DAPI, & cytoskeletal marker MAP2. (F) WT KOLF & Alsin-/- iPSC-sMN were immunostained with antibodies against Rab5 & Alsin, along with DAPI (nuclear) & phalloidin (actin) stains. Scale bars, 10 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29469808), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: MAP2 Antibody [NB300-213] - PEA decreases astrocyte activation in organotypic cultures of rat hippocampi & rescues neuronal CA3 damage caused by A beta challenge. A beta -challenged (1 μg/ml) slices of rat hippocampi treated for 24 hours w/ PEA (0.1 μM) in presence of selective PPAR gamma antagonist (GW9662, 9 nM) or selective PPAR alpha antagonist (MK886, 3 μM). (A) Nissl staining showing effect of treatment on morphology of organotypic hippocampal slices. Scale bar: 80 μm. A beta caused a marked neuronal loss, mainly in CA3 region of hippocampus, as highlighted by arrows. PEA able to reverse this effect. (B) Representative photomicrographs of CA3 region showing Results of immunofluorescence experiments aimed at investigating effect of treatments on astrocyte activation & neuronal loss, as determined by immunostaining for GFAP (green) & MAP2 (red) alone or merged, respectively. Nuclei stained w/ Hoechst (blue). Scale bar: 10 μm. Arrows in photomicrographs indicate astrocyte infiltration events & apoptotic condensation in nuclei of adjacent neurons. (C) Relative quantification of GFAP-positive cell number as a count of astrocyte proliferation. (D) Apoptotic events detected on MAP2-expressing cells as an indication of neuronal death. For (C) & (D), average value determined by counting cells in at least five microscopic fields for each treatment. Results are presented as means ± SEM of four separate experiments. At least four slices from each experimental group observed for each experiment. Statistical analysis performed using parametric one-way analysis of variance, & multiple comparisons performed using Bonferroni test. ***P < 0.001 & *P < 0.05 vs. control; ∘∘P < 0.01 & ∘P < 0.05 vs. A beta -challenge slices. Image collected & cropped by CiteAb from following publication (//jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094-9-49), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: MAP2 Antibody [NB300-213] - PEA inhibits astroglial proliferation & reduces neuronal loss in mixed neuroglia co-cultures exposed to A beta . A beta -challenged (1 μg/ml) astrocyte/neuron mixed cultures were treated with PEA (0.1 μM) in the presence of the selective PPAR gamma antagonist (GW9662, 9 nM) or the selective PPAR alpha antagonist (MK886, 3 μM). After 24 hours of treatment, cells were processed for analyses. (A) Immunofluorescence photomicrographs showing the effect of the treatments on astrocyte proliferation & neuronal loss, as determined by immunostaining for GFAP (green) & MAP2 (red), respectively. Arrows indicate chromatin condensation in nuclei stained with Hoechst dye (blue) as markers of apoptotic events. Scale bar: 10 μm. (B) Relative quantification of GFAP-positive cell number as a count of astrocyte proliferation. (C) Apoptotic events detected on MAP2-expressing cells as an indication of neuronal death. For (B) & (C), the average value was determined by counting cells in at least nine microscopic fields for each treatment. Results are presented as means ± SEM of three separate experiments. Statistical analysis was performed using parametric one-way analysis of variance, & multiple comparisons were performed using the Bonferroni test. ***P < 0.001 vs. unstimulated cells, ∘∘P < 0.01 & ∘P < 0.05 vs. A beta -stimulated cultures. Image collected & cropped by CiteAb from the following publication (//jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094-9-49), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Fe

Species Glossary

Reactivity: Hu, Mu, Rt, Fe

Applications:

, , ,

Applications: , , ,

Host:

Chicken Polyclonal

Host: Chicken Polyclonal

Conjugate:

Unconjugated

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NB100-309

ATM Antibody (2C1) - Azide and BSA Free

(3 Reviews)

(132 Publications)

100 ul

NB100-309

Western Blot: ATM Antibody (2C1) [NB100-309] - Analysis of ATM in U2OS sarcoma cells using ATM antibody (2C1) [NB100-309]. Theoretical molecular weight 351 kDa. Western blot image submitted by a verifeid customer review.

Immunohistochemistry-Paraffin: ATM Antibody (2C1) [NB100-309] - Human Kidney (formalin-fixed, paraffin-embedded) stained with ATM antibody (2C1) [NB100-309] at 5 ug/ml followed by biotinylated anti-mouse IgG secondary antibody, alkaline phosphatase-streptavidin and chromogen.

<b>Genetic Strategies Validation. </b>Western Blot: ATM Antibody (2C1) [NB100-309] - Non-transfected (-) and transfected (+) HeLa whole cell extracts (60 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with ATM antibody (2C1) [NB100-309] diluted at 1:500.

<b>Genetic Strategies Validation. </b>Western Blot: ATM Antibody (2C1) [NB100-309] - Methylated EDSBs may be repaired by an ATM-dependent pathway. Immunoblots of ATM and DNA-PKcs in ATM shRNA-transfected HeLa cells, using ATM antibody (2C1) [NB100-309]. GAPDH is included as a loading control. Image collected and cropped by CiteAb from the following publication (//molecular-cancer.biomedcentral.com/articles/10.1186/1476-4598-9-70), licensed under a CC-BY license.

Immunoprecipitation: ATM Antibody (2C1) [NB100-309] - MDA-MB-231 whole cell lysates were immunoprecipitated with 2 ug ATM Antibody (NB100-309), followed by Western blot (primary antibody: NB100-309 at 1:1000, 4C overnight. Western blot image submitted by a verified customer review.

<b>Biological Strategies Validation. </b>Western Blot: ATM Antibody (2C1) [NB100-309] - MCELNs decreased the DNA damage of H9C2 cells after radiation. Western blot images of p-ATM (NB100-306) and ATM (NB100-309) in H9C2 cells after 48 h of culture with indicated treatment. IR (-/+): 0/16 Gy X-ray; MCELNs (-/+): 0/10 ug/mL. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35509278/) licensed under a CC-BY license.

Western Blot: ATM Antibody (2C1) [NB100-309] - Western blot analysis of the extent of knockdown MRE11 & ATM, & the lack of effect of Mirin activation of ATM. The extent of shRNA-mediated knockdown of MRE11 & ATM are shown for (A) clones GFP-7F1 & GFP-6D1 & (B) clones EDS-7F2 & EDS-6J8. The extent of knockdown was determined by analyzing the intensity of the ATM & MRE11 bands relative to the intensity of the loading control GAPDH bands (see Table 1). (C) The effect of Mirin on activation of ATM was determined by analysis of phosphorylation of ATM in response to ionizing radiation. Cultures treated with DMSO alone, 20 μM Mirin, or knockdown of ATM were analyzed by western blot 30 min after exposure to 10 Gy of ionizing radiation. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26209132), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ATM Antibody (2C1) [NB100-309] - Rac1 inhibition abolishes IR-induced activation of both ATM & ATR signaling. (A) MCF-7 cells were treated with/without 20-Gy IR in the presence or absence of 100 μM NSC23766 & incubated for 1 hour at 37°C before analysis. To assess the ATM kinase activity, ATM was immunoprecipitated from cell lysate by using anti-ATM antibody (2C1) & assayed for ATM activity by using p53 recombinant protein as substrate. To measure the Chk2 activity, Chk2 was immunoprecipitated from cell lysate by using B-4 anti-Chk2 antibody & assayed for Chk2 activity by using Cdc25C recombinant protein as substrate. As controls, ATM & Chk2 protein levels in the immunoprecipitates (IP-WB) as well as in cell lysates (WB) were assessed with immunoblotting. (B) ATR & Chk1 were immunoprecipitated from cell lysates by using N-19 anti-ATR & G-4 anti-Chk1 antibody, respectively. ATR activity was assayed by using p53 recombinant protein substrate, & Chk1 activity assayed by using Cdc25C recombinant protein substrate. As controls, ATR & Chk1 protein levels in the immunoprecipitates (IP-WB), as well as in cell lysates (WB) were assessed with immunoblotting. (C) MCF-7 cells were exposed to IR at the indicated doses in the presence or absence of NSC23766, incubated for 1 hour, & assessed for Chk1 & Chk2 activities. *Kinase assay does not contain Cdc25C substrate. (D) T47D & ZR-75-1 cells were exposed to 10-Gy IR in the presence or absence of 100 μM NSC23766, incubated for 1 hour, & analyzed for Chk1 & Chk2 activities. Image collected & cropped by CiteAb from the following publication (//breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3164), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ATM Antibody (2C1) [NB100-309] - Inhibition of Rac1 by N17Rac1 mutant or Rac1 siRNA diminishes IR-induced G2/M checkpoint activation. (A) MCF-7 cells were infected with Ad.N17Rac1 or Ad.Control for 24 hours & exposed to 15-Gy IR. Left panel: the cells were analyzed for DNA content 24 hours after IR. The result depicts the percentage of cells with 4N-DNA content & is shown as mean ± SD of quadruplicate samples. *P < 0.001 (n = 4), significant difference from the irradiated Ad.Control-infected cells. Right panel: Inset: at 15 minutes after IR, the infected cells were analyzed for Rac1 activities (Rac1-GTP) & protein levels (total Rac1). Bar graph: mitotic cells in the cell samples were analyzed 2 hours after IR. The result depicts the percentage of mitotic cells & is shown as mean ± SD of triplicate samples. **P = 0.002 (n = 3), significant difference from the irradiated Ad.Control-infected cells. (B) Upper panel: MCF-7 cells transfected with Rac1 siRNA (Rac1) or control siRNA (Control) were incubated for the indicated times & analyzed for protein levels of Rac1 & Actin. Lower panel: After 2-day incubation, the siRNA-transfected cells were exposed to IR, incubated for 24 hours, & assessed for DNA content. Results depict the percentage of cells with 4N-DNA content & represent the mean ± SD of three separate experiments in duplicate samples. *P < 0.001 (n = 6), significant difference from the irradiated Control-siRNA transfected cells. (C) After 2-day incubation, siRNA-transfected cells were treated with/without 20-Gy IR, incubated for 1 hour, & analyzed for ATM, ATR, Chk1, & Chk2 activities. Image collected & cropped by CiteAb from the following publication (//breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3164), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ATM Antibody (2C1) [NB100-309] - HeLa whole cell extract and nuclear extracts (30 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with ATM antibody [2C1] (NB100-309) diluted at 1:500. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.

Western Blot: ATM Antibody (2C1) [NB100-309] - Whole cell extract (30 ug) was separated by 5% SDS-PAGE, and the membrane was blotted with ATM antibody [2C1] (NB100-309) diluted at 1:1000.

Human Testis (formalin-fixed, paraffin-embedded) stained with ATM antibody at 5 ug/ml followed by biotinylated anti-mouse IgG secondary antibody, alkaline phosphatase-streptavidin and chromogen.

Reactivity:

Hu, Mu, Rt, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, Pm

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Conjugate:

Unconjugated

Validated by:

Validated by:

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NB300-141

GFAP Antibody

(12 Reviews)

(117 Publications)

0.05 ml

NB300-141

Simple Western: GFAP Antibody [NB300-141] - Simple Western lane view shows a specific band for GFAP in 0.05 mg/mL of Human Brain lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Immunocytochemistry/Immunofluorescence: GFAP Antibody [NB300-141] - The inhibition of Kv1.3 channels induces neuroprotection against the toxic effects of glioma. Immunofluorescence analyses of Kv1.3 expression (red) on GFAP-positive cells (green) in coronal brain slices of GL261-bearing mice in peritumoral region; in the magnification (right) arrows indicate co-expression of Kv1.3 and GFAP in the same cell. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-018-25940-5) licensed under a CC-BY license.

Immunohistochemistry-Paraffin: GFAP Antibody [NB300-141] - Mouse brain section, 20x magnification. Antibody at 1:1000. Detection with Polymer-HRP. IHC image submitted by a verified customer review.

Western Blot: GFAP Antibody [NB300-141] - Analysis of Rat brain lysate. Antibody at 1:5000. Specific band running with an apparent SDS-PAGE molecular weight of ~50 kDa corresponds to rodent GFAP was observed.

Western Blot: GFAP Antibody [NB300-141] - Analysis of GFAP expression in whole rat cerebellum homogenate.

Western Blot: GFAP Antibody [NB300-141] - Analysis of different tissue lysates using rabbit polyclonal antibody to GFAP, NB300-141, dilution 1:5000 in green: [1] protein standard (red), [2] rat brain, [3] rat spinal cord, [4] mouse brain, [5] mouse spinal cord. Strong band at about 50 kDa corresponds to the major isotype of the GFAP protein. Smaller isotypes and proteolytic fragments of GFAP are also detected on the blot.

Immunocytochemistry/Immunofluorescence: GFAP Antibody [NB300-141] - Analysis of mixed neuron-glial cultures using GFAP antibody NB300-141 (red) and Vimentin antibody NB300-223 (green). The fibroblastic cells contain only Vimentin and so are green. The astrocytes contain either Vimentin and GFAP (appearing golden) or predominantly GFAP (appearing red). Blue is nuclear DNA stain.

Immunocytochemistry/Immunofluorescence: GFAP Antibody [NB300-141] - Cultured Rat hippocampal neurons. ICC/IF image submitted by a verified customer review.

Immunohistochemistry: GFAP Antibody [NB300-141] - Xenografted mouse brain section: astocyte and human nuclei. ICC/IF image submitted by a verified customer review.

Immunohistochemistry-Frozen: GFAP Antibody [NB300-141] - Imaging of mouse brain (cortex), 20x magnification. IHC image submitted by a verified customer review.

EGFP+ neurons are positive for RABV antigen.Brains were collected from Cre reporter mice fifteen days post-infection, cryosectioned, and EGFP+ regions compared to cell-specific labeling, A) NeuN (blue, neuronal nuclei antibody, 20× fluorescence imaging), B) GFAP (blue, astrocyte antibody, 40× confocal imaging), or C) RABV P antigen (purple) and DAPI nuclear stain (blue, 63× confocal imaging). White arrows in (C) indicate regions positive for RABV P.

Immunocytochemistry/ Immunofluorescence: GFAP Antibody [NB300-141] - EGFP+ neurons are positive for RABV antigen.Brains were collected from Cre reporter mice fifteen days post-infection, cryosectioned, & EGFP+ regions compared to cell-specific labeling, A) NeuN (blue, neuronal nuclei antibody, 20× fluorescence imaging), B) GFAP (blue, astrocyte antibody, 40× confocal imaging), or C) RABV P antigen (purple) & DAPI nuclear stain (blue, 63× confocal imaging). White arrows in (C) indicate regions positive for RABV P. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.ppat.1002971), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: GFAP Antibody [NB300-141] - Effects of the repeated administration of NVP CXCR2 20 (NVP; 10 μg/5 μl; i.t.; 16 h & 1 h before CCI & then once a day for 7 days) on the protein levels of CXCR2, IBA1, GFAP, CXCL1, CXCL2, & CXCL3 proteins (A–I) in the spinal cord (A–F) & DRG (G–I) on the 7th day after CCI in rats. The data are presented as the mean fold changes relative to the control ± SEM (5–6 samples per group). Intergroup differences were analyzed using ANOVA with Bonferroni's multiple comparisons test. *p < 0.05, **p < 0.01, ***p < 0.001 indicate differences vs. naive rats. #p < 0.05, indicate differences between V-treated & NVP-treated rats. CCI, chronic constriction injury; N, naive; V, vehicle; NVP, NVP CXCR2 20. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31616413), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: GFAP Antibody [NB300-141] - Increased astrocyte activation in IL-1 beta -stimulated two-week old offspring is reduced by maternal LB supplementation. Representative images of fluorescence microscopy of claudin-5+ (location of the brain capillaries, red), GFAP+ astrocyte (green), & DAPI (nuclei, blue). Three to five sections per mouse were examined & at least three mice were examined in each group. Stronger than control SPF GFAP staining was observed around the blood vessel (see arrow) after IL-1 beta insult. Maternal supplemented group (LB) with or without postnatal insult had GFAP levels similar to the control group indicating that LB supplementation prevented astrocyte activation around the BBB endothelium. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32424168), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: GFAP Antibody [NB300-141] - Glial cells density is differently modulated during AD progression.a Retinal slices were immunolabeled with anti-GFAP antibody (green) & Hoechst for nuclei visualization (blue) at different ages of 3xTg-AD & non-Tg mice & density of GFAP signal was quantified as shown in b (**p < 0.01 pre vs early; n = 16 fields/four slices for each condition; two-way ANOVA, Holm-Sidak; ##p < 0.01 for comparison with age-matched non-Tg mice, two-way ANOVA, Holm-Sidak). c Representative multiarea image of retinal slice immunolabeled with anti-Iba1 antibody (green) & Hoechst for nuclei visualization (blue); density of Iba1+ cells was quantified as shown in d (**p < 0.01 vs 3xTg-AD pre; ##p < 0.01 vs age-matched non-Tg mice, n = 16 fields/four slices for each condition, two-way ANOVA, Holm-Sidak method for multiple comparison) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29880901), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Bv, Ch, Eq, Gp, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Ch, Eq, Gp, Rb

Applications:

, , ,

Applications: , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

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NB600-633

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

(4 Reviews)

(116 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.

Western Blot: IL-1 beta/IL-1F2 Antibody [NB600-633] - Methylene blue inhibits inflammasome formation in cultured microglia. (A) Expression of Pro-Caspase-1, Caspase-1 p20, pro-IL-1 beta & mature IL-1 beta in microglia with or without stimulation in the presence or absence of methylene blue. Left: representative Western blot images. Right: statistics. N = 5 per group. The (B) expression of NLRP3, NLRC4 & Aim2 in microglia. Left: representative Western blot images. Right: statistics. N = 4 per group. (C) Co-immunoprecipitation assay showing the binding of ASC to NLRP3 or NLRC4. Left: representative Western blot images. Right: statistics. Un: no stimulation. LPS+ATP: stimulation with LPS followed by ATP treatment. V: vehicle; M: 500 nM methylene blue. IP: immunoprecipitation with the antibody against indicated protein. IB: detection of indicated protein. To analyze the relative expression of target proteins, the band intensities of target proteins were normalized to corresponding band intensities of alpha -Tubulin, followed by calculating the expression in other groups relative to “Un (V)” group. N = 4 per group. *p < 0.05; **p < 0.01; ***p < 0.001 in comparison with unstimulated cells of the vehicle group. #p < 0.05; ##p < 0.01 in comparison with stimulated cells of the vehicle group. Image collected & cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fncel.2017.00391/full), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Rabbit Polyclonal IL-1 beta/IL-1F2 Antibody [NB600-633] - IL-1 beta/IL-1F2 Western blot of liver homogenate from control (1) and alcohol-fed mice (2). Image from a verified customer review.

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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NB600-363

HA Tag Antibody - BSA Free

(3 Reviews)

(111 Publications)

0.1 ml

NB600-363

Western Blot: HA Tag Antibody [NB600-363] - Wnt/beta-catenin signaling regulates HySp5 expression. Immunoprecipitation (IP) of HA-tagged HySp5-420 expressed in HEK293T cells together or not with hudeltabeta-Catenin (upper) or huTCF1 (lower). IP was performed with an anti-HA antibody and Co-IP products were detected with the anti-beta-catenin or anti-TCF1 antibodies. Same results were obtained in two independent experiments. Each data point represents one biological independent experiment. Statistical p values: *p <= 0.05, **p <= 0.01, ***p <= 0.001, ****p <= 0.0001 (unpaired t test). Error bars indicate SD. PP primer pair, RLA relative luciferase activity Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-018-08242-2), licensed under a CC-BY license.

Western Blot: HA Tag Antibody [NB600-363] - ZFN expression levels. After co-transfection of HEK293T cells with ZFN expression vectors and pEGFP, cell lysates were probed with antibodies against the HA tag and EGFP. Amount of transfected ZFN plasmids was 75 ng, 300 ng, and 1200 ng. NT, non-transfected cells. Selection-independent generation of gene knockout mouse embryonic stem cells using zinc-finger nucleases. PLoS One (2011)

Immunocytochemistry/Immunofluorescence: HA Tag Antibody [NB600-363] - HA-tagged proteins were detected in immersion fixed HEK293 human embryonic kidney cell line transfected with HA-tagged MGAT2 using 1 ug/mL goat anti-HA Tag polyclonal (NB600-362, Novus Biologicals), 1 ug/mL rabbit anti-HA Tag polyclonal (NB600-363, Novus Biologicals). Cells were stained using the appropriate secondary antibody, donkey anti-mouse IgG-NL557 (NL007) and counterstained with DAPI (blue).

Western Blot: HA Tag Antibody [NB600-363] - Gene knockout in U2OS.693 cells.(A) Schematic of ZFN-mediated knockout. A ZFN pair (ZFNR & ZFNL) designed to target position 502 in the EGFP gene (E502) creates a DNA double-strand break that is sealed by the error-prone non-homologous end-joining (NHEJ) pathway & hence leads to disruption of the coding sequence. (B) Dose-dependent gene disruption in U2OS.693 cells. U2OS.693 cells that stably express a destabilized EGFP were transfected with increasing amounts of E502-specific ZFN expression vectors (75–1200 ng). The percentage of EGFP-negative cells was determined 6 days post-transfection by flow cytometry (n = 3; indicated is average & standard deviation). E502-WT, ZFN with wild-type FokI domain; E502-OH, ZFN with obligate heterodimeric FokI domain. (C) ZFN expression levels. After co-transfection of HEK293T cells with ZFN expression vectors & pEGFP, cell lysates were probed with antibodies against the HA tag & EGFP. Amount of transfected ZFN plasmids was 75 ng, 300 ng, & 1200 ng. NT, non-transfected cells. (D) Kinetics of EGFP knockout. The graph displays the percentage of EGFP-negative cells (see B) from day 1 to day 6 post-transfection for two vector amounts (n = 3; indicated is average & standard deviation). WT, E502-WT; OH, E502-OH. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/22194948), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

ET

Species Glossary

Reactivity: ET

Applications:

, , , , , , , ,

Applications: , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB600-363AF350
Alexa Fluor 405	NB600-363AF405-0.1ml
Alexa Fluor 488	NB600-363AF488-0.1ml
Alexa Fluor 532	NB600-363AF532
Alexa Fluor 594	NB600-363AF594
Alexa Fluor 647	NB600-363AF647-0.1ml
Alexa Fluor 700	NB600-363AF700-0.1ml
Alexa Fluor 750	NB600-363AF750
Allophycocyanin	NB600-363APC
Biotin	NB600-363B
CoraFluor 1	NB600-363CL1
DyLight 350	NB600-363UV-0.1ml
DyLight 405	NB600-363V-0.1ml
DyLight 488	NB600-363G-0.1ml
DyLight 550	NB600-363R-0.1ml
DyLight 594	NB600-363DL594
DyLight 650	NB600-363C-0.1ml
DyLight 680	NB600-363FR-0.1ml
DyLight 755	NB600-363IR-0.1ml
FITC	NB600-363F
HRP	NB600-363H-0.1ml
Janelia Fluor 525	NB600-363JF525
Janelia Fluor 549	NB600-363JF549
Janelia Fluor 646	NB600-363JF646
Janelia Fluor 669	NB600-363JF669
PE	NB600-363PE
PerCP	NB600-363PCP
mFluor Violet 450 SE	NB600-363MFV450
mFluor Violet 500 SE	NB600-363MFV500
mFluor Violet 610 SE	NB600-363MFV610

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2275-PC-100

GAPDH Antibody [Unconjugated]

(111 Publications)

100 uL

2275-PC-100

Western blot shows lysates of U2OS human osteosarcoma cell line, A20 mouse B cell lymphoma cell line, and L6 rat myoblast cell line. PVDF membrane was probed with 0.1 µg/mL Rabbit Anti-Human/Mouse/Rat GAPDH Polyclonal Antibody (Catalog # 2275-PC) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for GAPDH at approximately 36 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western shows lysates of Exosome Standards (LNCaP) (NBP3-11687), Exosome Standards (HT‑29) (NBP3-11685) and Jurkat human acute T cell leukemia cell line, loaded at 0.5 mg/ml. A specific band was detected for GAPDH at approximately 41 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse/Rat GAPDH Antigen Affinity-purified Polyclonal Antibody (Catalog # 2275-PC-100). This experiment was conducted under reducing conditions and using the 12-230kDa separation system.

Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line, C2C12 mouse myoblast cell line, and C6 rat glioma cell line, loaded at 0.2 mg/mL. A specific band was detected for GAPDH at approximately 43 kDa (as indicated) using 1 µg/mL of Rabbit Anti-Human/Mouse/Rat GAPDH Antigen Affinity-purified Polyclonal Antibody (Catalog # 2275-PC-100) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Higher levels of CD4 are found in macrophages expressing LCK.A) Schematic diagram of self-inactivating lentiviral construct used to express wild-type Lck and puroR (LCKWT P). A similar construct was used to express a kinase inactive form of Lck and puroR (LCKINACTIVE P) or to express puroR only (‘control P’). B) Detection of protein expression of LCK and CD4. Control and transgenic PSC-macrophages lysates were analysed by western blotting using anti-LCK and anti-CD4 antibodies. The loading control GAPDH was detected using anti-GAPDH antibody. C) Protein levels were measured with Odyssey software (Li-COR) and CD4 expression was normalised to GAPDH expression. Symbols represent normalised CD4 expression, relative to the PSC-macrophages control group, of two independent experiments. D) Detection of surface CD4 and total LCK expression. Representative two-colour immunofluorescence (dot plot) analysis is shown. Gates were determined by using the two relevant isotype control antibodies. Quantification of total LCK expression (E) and surface CD4 expression (F), expressed as the ratio of the geometric mean fluorescence intensity (MFI) over the isotype control ±SEM of independent experiments (n = 7). G) Detergent resistance of CD4, expressed as mean Flow Cytometric Detergent Resistance (FCDR) index of CD4 (n = 4) in PSC-macrophages, calculated as described in materials and methods. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24465876), licensed under a CC-BY license. Not internally tested by R&D Systems.

CD4 knock-down in genetically modified stem cell-derived macrophages. C) Detection of protein expression of CD4. Control and transgenic PSC-macrophages lysates were analysed by western blotting using anti-CD4 antibodies. GAPDH, a loading control, was detected using anti-GAPDH antibody. Representative blot is shown. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24465876), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, ,

Applications: , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

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NBP1-49461

Stathmin-2/STMN2 Antibody - BSA Free

(2 Reviews)

(102 Publications)

0.1 mg

NBP1-49461

<b>Genetic Strategies Validation. </b>Staining of STMN2 in primary mouse dorsal root ganglia (DRG) neurons Image shows the expected staining of endogenous STMN2 in axons and growth cones. Tubulin is a marker of axons. NBP1-49461 was used at a dilution of 1:4000. Image courtesy of Dr. Jung Eun Shin.

Neuronal deletion of Tsc2 improves axon regeneration within the first 2 d following nerve injury. SCG10 immunostaining of sciatic nerves 1 d after crush injury in control and Tsc2 cKO mice with (conditioned) or without (sham) a conditioning injury. Red dotted line denotes the crush site while red arrowheads point to the three longest axons. Scale bar: 500 i1/4m. Image collected and cropped by Citeab from the following publication (Nociceptor Deletion of Tsc2 Enhances Axon Regeneration by Inducing a Conditioning Injury Response in Dorsal Root Ganglia. Eneuro (2019) licensed under a CC-BY license.

<b>Expression of Tagged Proteins Validation. </b>Staining of STMN2 in HEK293T cells transfected with GFP or a GFP-STMN2 fusion. NBP1-49461 was used at a dilution of 1:1000 (A, B) and 1:3000 (C). Image courtesy of Dr. Jung Eun Shin.

<b>Expression of Tagged Proteins Validation. </b>Analysis of STMN2 in HEK293T cells transfected with GFP or a GFP-STMN2 fusion. NBP1-49461 was used at a dilution of 1:4000. Image courtesy of Dr. Jung Eun Shin.

Reactivity:

Hu, Mu, Rt, Bv

Species Glossary

Reactivity: Hu, Mu, Rt, Bv

Applications:

, , , , ,

Applications: , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP1-49461AF350
Alexa Fluor 405	NBP1-49461AF405
Alexa Fluor 488	NBP1-49461AF488
Alexa Fluor 532	NBP1-49461AF532
Alexa Fluor 594	NBP1-49461AF594
Alexa Fluor 647	NBP1-49461AF647
Alexa Fluor 700	NBP1-49461AF700
Alexa Fluor 750	NBP1-49461AF750
Allophycocyanin	NBP1-49461APC
Biotin	NBP1-49461B
CoraFluor 1	NBP1-49461CL1
DyLight 350	NBP1-49461UV
DyLight 405	NBP1-49461V
DyLight 488	NBP1-49461G
DyLight 550	NBP1-49461R
DyLight 594	NBP1-49461DL594
DyLight 650	NBP1-49461C
DyLight 680	NBP1-49461FR
DyLight 755	NBP1-49461IR
FITC	NBP1-49461F
HRP	NBP1-49461H
Janelia Fluor 525	NBP1-49461JF525
Janelia Fluor 549	NBP1-49461JF549
Janelia Fluor 646	NBP1-49461JF646
Janelia Fluor 669	NBP1-49461JF669
PE	NBP1-49461PE
PerCP	NBP1-49461PCP
mFluor Violet 450 SE	NBP1-49461MFV450
mFluor Violet 500 SE	NBP1-49461MFV500
mFluor Violet 610 SE	NBP1-49461MFV610

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NBP1-05987

ZEB1 Antibody

(6 Reviews)

(102 Publications)

0.1 ml

NBP1-05987

Western Blot: ZEB1 Antibody [NBP1-05987] - Analysis of ZEB1 in transfected and native mouse cell lysate. Image courtesy of anonymous customer review.

Immunohistochemistry-Paraffin: ZEB1 Antibody [NBP1-05987] - Analysis of ZEB1 in human liver and spleen tissues. Image courtesy of anonymous customer review.

<b>Independent Antibodies Validation. </b>Immunoprecipitation: ZEB1 Antibody [NBP1-05987] - Detection of human ZEB1 by western blot of immunoprecipitates. Samples: Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from HeLa cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-ZEB1 antibody NBP1-05987 used for IP at 3 ug per reaction. ZEB1 was also immunoprecipitated by another rabbit anti-ZEB1 antibody. For blotting immunoprecipitated ZEB1, NBP1-05987 was used at 0.4 ug/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.

Immunohistochemical staining results of EMT regulators in HG‐PanIN: SNAIL, SLUG, TWIST1, and ZEB1. We categorized and defined three grades: grade 0, <10% positive staining; grade 1, 10%‐50% positive; grade 2, 50%< positive

Western Blot: ZEB1 Antibody [NBP1-05987] - Lin28a & ETM-related protein expression changed after the knockdown of Zeb1 in mESCs, mEB, GC1, & P19. Western blotting demonstrated that E-cadherin protein expression increased & that Lin28a & Vimentin protein expression decreased in Zeb1-siRNA transfected mESC (panel a), mEB (panel b), GC1 (panel c), & P19 (panel d) cells. Each experiment was performed in triplicate. Abbreviations: NC, negative control; siRNA, small interfering RNA; & WT, wild-type. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35611144), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: ZEB1 Antibody [NBP1-05987] - Immunohistochemical staining results of EMT regulators in HG‐PanIN: SNAIL, SLUG, TWIST1, & ZEB1. We categorized & defined three grades: grade 0, <10% positive staining; grade 1, 10%‐50% positive; grade 2, 50%< positive Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30791220), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: ZEB1 Antibody [NBP1-05987] - Location of Zeb1 proteins in mEC lines (mESC, mEB, GC1, & P19). Immunofluorescence analysis was used to determine localization of Zeb1 in mESC, mEB, GC1, & P19 using a Zeb1 antibody (red). Cell nuclei were labeled with Hoechst 33342 (blue). The scale for the measurement bar is 50 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35611144), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ZEB1 Antibody [NBP1-05987] - Zeb1 expression in mESCs, mEB, GC1, & P19. (a) Zeb1 mRNA expression detected by real-time qPCR in mESC, mEB, GC1, & P19 (P < 0.001, data presented as mean value ± SD). (b) Zeb1 protein expression examined by western blot in mESC, mEB, GC1, & P19. Each experiment was performed in triplicate. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35611144), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: ZEB1 Antibody [NBP1-05987] - Correlation of the expression of ZEB1 & IL‐6 & other cytokines in various cancers. (A) Correlation analysis of the expression of ZEB1 & cytokines in breast cancer cell lines using data obtained from the Cancer Cell Line Encyclopedia (CCLE). Breast cancer cell lines were selected for the analysis, & each dot represents a cell line. Affymetrix microarray probe IDs for IL6 & IL8 are shown after the gene symbols on the y‐axis. RMA, robust multiarray average. (B) Representative results obtained from the tissue array analysis in Fig. S3. The top panel shows a case that was positive for both nuclear ZEB1 staining (green) & whole‐cell IL‐6 staining (red); scores = 4. The bottom panel shows a case that was negative for both ZEB1 & IL‐6; scores = 1. The samples were counterstained with DAPI to show cell density in the spot. Original magnification: 20 ×. (C) Kaplan–Meier survival curves of breast & lung cancer patients obtained from a public meta‐analysis database & Kaplan–Meier plotter (Gyorffy et al., 2010, 2013). The probability of overall survival of patients as split by median IL6 & IL8 expression is shown. Red: IL6 & IL8 high expression group; black: IL6 & IL8 low expression group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28618162), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ZEB1 Antibody [NBP1-05987] - SU5402, FGFR1 inhibitor, affects the EMT transcription factors.(A, B) ERK1/2 phosphorylation (p-ERK1/2) determined by immunoblotting in HSC-4 & OTC-04 treated for 30 min w/ 30 ng/ml FGF2 or 30 ng/ml FGF7 in presence of 10% FBS (A) & in TSU & HOC313 cells treated for 1 h w/ 30 ng/ml FGF2 or 30 μM SU5402 in absence of FBS (B). F2, FGF2; F7, FGF7; SU, SU5402. (C) We have previously reported that, after treatment w/ TGF-beta , NMuMG cells underwent EMT w/ the IIIc-isoform of FGFR1[17]. After NMuMG cells pretreated w/ TGF-beta transfected w/ mouse Fgfr1 siRNA or treated w/ SU5402, the cells further incubated in culture medium (CM) from TSU cells. SU, SU5402; siFR1, siRNA against mouse Fgfr1. (D)FGF2 mRNA levels determined by RT-qPCR analyses. The ratio of FGF2 mRNA to GAPDH mRNA in HSC-4 cells indicated as “1”. Each value represents the mean ± SD of triplicate determinations from a representative experiment. Similar results obtained in at least three independent experiments. (E) ERK1/2 phosphorylation (p-ERK1/2) in TSU & HOC313 cells monitored in presence of indicated concentration of SU5402 for 1 h under serum-free culture conditions, followed by immunoblot analysis. (F, G) Expression of indicated genes in TSU cells under serum-free culture conditions determined by RT-qPCR (F) & immunoblot (G) analyses, following treatment w/ 10 μM SU5402. Each value represents the mean ± SD of triplicate determinations from a representative experiment. Similar results obtained from at least three independent experiments. p values determined by Student’s t-test. **p < 0.01. (H) TSU cells treated w/ 10 μM U0126 in absence of FBS subjected to immunoblotting w/ the indicated antibodies. alpha -tubulin used as a loading control (A, B, C, E, G, & H). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31682640), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ZEB1 Antibody [NBP1-05987] - Vimentin & ZEB1 protein levels are regulated by OVOL1 & OVOL2 in a human SCC cell line. (A) Relative protein expression levels of OVOL1 & OVOL2 in A431 cells treated with control siRNA, OVOL1 siRNA, or OVOL2 siRNA for 48 h. (Left) Representative blot images & (right) relative expression levels calculated from three independent experiments. (B) Relative protein expression levels of vimentin, ZEB1, & E-cadherin in A431 cells treated with control siRNA, OVOL1 siRNA, or OVOL2 siRNA for 72 h. (Upper) representative blot images & (lower) relative protein expression levels calculated from three independent experiments. Mann–Whitney U-test; error bars represent mean ± standard deviation. Protein expressions are relative to those of beta -actin as a reference. The control siRNA value was set to 1. p-values < 0.05 were assumed to indicate a statistically significant difference; * p < 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32106476), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ZEB1 Antibody [NBP1-05987] - PDGFR-alpha / beta promotes sarcoma CSC migration/invasion & anchorage-independent growth.a Western blot analysis of cell adhesion protein N-cadherin & EMT transcription factors Snail, Slug, & Zeb1 in human sarcoma cell lines grown as monolayers & as spheroids. beta -actin was used as the loading control. b Representative images under light microscopy of migration & invasion assays of human sarcoma cell lines grown as monolayers or as spheroids for 24–48 h. Graphs display the number of migrated or invasive cells per field. c Representative light microscopy images of forrmed colonies in soft agar assay of human sarcoma cell lines grown as monolayers or spheroids for 14–20 days. Graph displays the number colonies per field. d Representative images under light microscopy of migration & invasion assays of human sarcoma cell lines grown as monolayers or as spheroids. Spheroid cells were treated with imatinib or DMSO. e Western blot analysis of N-cadherin & EMT-regulating transcription factors in human sarcoma cell lines grown as spheroids & treated with imatinib or DMSO. Experiments in a & e were performed three times with similar results. Bars represent standard deviation. *p < 0.05 compared to Monolayers Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41389-018-0059-1), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ZEB1 Antibody [NBP1-05987] - FGFR1 siRNAs attenuate the malignant phenotypes of cancer cells.(A) mRNA from TSU cells transfected with siRNAs against FGFR1 (siFGFR1) were subjected to conventional RT-PCR to determine the levels of endogenous FGFR1. (B, C, D) After transfection with siFGFR1 in TSU cells, phosphorylation of ERK1/2 (p-ERK1/2) (B), ZEB1 levels (C), & cell morphology (D) were determined. (E) TSU cells transfected with siFGFR1 were subjected to immunofluorescence analyses. Low magnification, 40×; high magnification, 80×. (F, G, H, I) After either treatment with SU5402 or transfection with siFGFR1 in TSU & HOC313 cells, the number of cells & invasive properties were determined under serum-free culture conditions. (J & K) After transfection with siFGFR1c in TSU & HOC313 cells, invasive & migratory properties were determined under serum-free culture conditions in TSU & HOC313 cells, respectively. siNC, non-specific control siRNA. Each value represents the mean ± SD of triplicate determinations from a representative experiment. Similar results were obtained from at least three independent experiments. p values were determined by Student’s t-test. *p < 0.01. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31682640), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ZEB1 Antibody [NBP1-05987] - Nav1.4 protein levels are decreased in muscles from mouse models of SMA. (A) Immunoblot analysis using muscle lysate from P2, P5, P9, & P21 wild type mice. Nav1.4 protein levels increase during postnatal muscle development & form the predominant sodium channel expressed in mature skeletal muscle. GAPDH served as a loading control (N = 3). (B) Representative immunoblot with quantification, showing a decrease in levels of sodium channel Nav1.4 & Nav1.5 in P5 Smn-/-;SMN2 hindlimb muscle compared with controls (N = 3). (C) Quantification of immunoblot analyses in P21 Smn2B/- & control hindlimb muscles revealed a decrease in Nav1.4 levels. Early in postnatal muscle development, the Nav1.5 sodium channel isoform is the most predominant. In P21 Smn2B/- mice, the protein levels of Nav1.5 are higher than in controls (N = 3). (D) The protein level of the Nav1.4 positive regulator, NF1, is not altered in muscles from P21 Smn2B/- mice. Similarly, no change was detected in the protein levels of the Nav1.4 repressor ZEB. (E) Expression of sodium channel Nav1.4 in control sham & denervated samples 1 & 7 days post-denervation was assessed by immunoblot (N = 3). A decrease in the levels of Nav1.4 in muscle was noted at 7 days post-denervation. *, P < 0.05; **, P < 0.01. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24119341), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

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Host: Rabbit Polyclonal

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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

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Antibody Validation

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Species Glossary

p62/SQSTM1 Antibody (2C11) - Azide and BSA Free

TDP-43/TARDBP Antibody (2E2-D3) - Azide and BSA Free

Histone H2AX [p Ser139] Antibody - BSA Free

CD31/PECAM-1 Antibody

MAP2 Antibody - BSA Free

ATM Antibody (2C1) - Azide and BSA Free

GFAP Antibody

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

HA Tag Antibody - BSA Free

GAPDH Antibody [Unconjugated]

Stathmin-2/STMN2 Antibody - BSA Free

ZEB1 Antibody