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![Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - WB analysis of recombinant p62 protein (immunogen, 74.51 KDa) using SQSTM1/p62 antibody (clone 2C11).](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-2C11-Western-Blot-H00008878-M01-img0012.jpg "Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - WB analysis of recombinant p62 protein (immunogen, 74.51 KDa) using SQSTM1/p62 antibody (clone 2C11).")
![<b>Genetic Strategies Validation. </b>Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - WB analysis of p62 in p62 WT and KO mouse hepatocytes and HepG2 cell lysate using anti-p62/SQSTM1 antibody clone 2C11. Sample lane1: p62 +/- mouse hepatocytes, lane 2: p62 -/- mouse hepatocytes, lane3: Wt mouse hepatocytes, lane 4: HepG2 cells. This image is from a verified customer product review.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-2C11-Western-Blot-H00008878-M01-img0005.jpg)
![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.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-2C11-Immunocytochemistry-Immunofluorescence-H00008878-M01-img0008.jpg)
![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.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-2C11-Immunocytochemistry-Immunofluorescence-H00008878-M01-img0010.jpg)
![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.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-2C11-Western-Blot-H00008878-M01-img0014.jpg)
![Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Analysis of p62/SQSTM1 antibody on HeLa cells. Image from verified customer review.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-2C11-Immunocytochemistry-Immunofluorescence-H00008878-M01-img0016.jpg)
![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.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-2C11-Western-Blot-H00008878-M01-img0015.jpg)
![ELISA: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Detection limit for recombinant GST tagged SQSTM1 is approximately 0.03ng/ml as a capture antibody.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-2C11-ELISA-H00008878-M01-img0013.jpg)
![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.](http://images.novusbio.com/fullsize/h00008878-m01_mouse-monoclonal-p62-sqstm1-antibody-2c11-31020241534382.jpg)
![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.](http://images.novusbio.com/fullsize/h00008878-m01_mouse-monoclonal-p62-sqstm1-antibody-2c11-310202415392568.jpg)
![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.](http://images.novusbio.com/fullsize/h00008878-m01_mouse-monoclonal-p62-sqstm1-antibody-2c11-310202415345313.jpg)
![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.](http://images.novusbio.com/fullsize/h00008878-m01_mouse-monoclonal-p62-sqstm1-antibody-2c11-310202415482234.jpg)
![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.](http://images.novusbio.com/fullsize/h00008878-m01_mouse-monoclonal-p62-sqstm1-antibody-2c11-310202415483626.jpg)
![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.](http://images.novusbio.com/fullsize/h00008878-m01_mouse-monoclonal-p62-sqstm1-antibody-2c11-31020241548511.jpg)
![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.](http://images.novusbio.com/fullsize/h00008878-m01_mouse-monoclonal-p62-sqstm1-antibody-2c11-310202415482212.jpg)
![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.](http://images.novusbio.com/fullsize/h00008878-m01_mouse-monoclonal-p62-sqstm1-antibody-2c11-31020241548515.jpg)
![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.](http://images.novusbio.com/fullsize/h00008878-m01_mouse-monoclonal-p62-sqstm1-antibody-2c11-310202415485130.jpg)
![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.](http://images.novusbio.com/fullsize/h00008878-m01_mouse-monoclonal-p62-sqstm1-antibody-2c11-310202415482274.jpg)
![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.](http://images.novusbio.com/fullsize/h00008878-m01_mouse-monoclonal-p62-sqstm1-antibody-2c11-310202415483640.jpg)
![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.](http://images.novusbio.com/fullsize/h00008878-m01_mouse-monoclonal-p62-sqstm1-antibody-2c11-310202415482248.jpg)
![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.](http://images.novusbio.com/fullsize/h00008878-m01_mouse-monoclonal-p62-sqstm1-antibody-2c11-31020241548226.jpg)
![Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - p62/SQSTM1 Antibody [NBP1-48320] - HeLa cells were treated overnight with 50uM CQ, then fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-p628/SQSTM1 at a 1:200 dilution overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Immunocytochemistry-Immunofluorescence-NBP1-48320-img0008.jpg "<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - p62/SQSTM1 Antibody [NBP1-48320] - HeLa cells were treated overnight with 50uM CQ, then fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-p628/SQSTM1 at a 1:200 dilution overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.")
![<b>Biological Strategies Validation. </b>Simple Western: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - p62/SQSTM1 Antibody [NBP1-48320] - Lane view shows a specific band for p62/SQSTM1 in 1.0 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Simple-Western-NBP1-48320-img0005.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - p62/SQSTM1 Antibody [NBP1-48320] - Cultured HeLa cells were treated with or without 50 uM chloriquine for 24 hours as indicated. Cell lysates were prepared and separated on a 12% gel by SDS-PAGE. Protein was transferred to PVDF membrane and blocked in 5% non-fat milk. The membrane was then probed with 1 ug/ml anti-p62/SQSMT1 in 1% milk and dectected with an anti-rabbit HRP secondary antibody using chemiluminescence. Note the upregulation of p62 (arrowhead) in response to chloroquine treatment and the blockage of autophagy.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Western-Blot-NBP1-48320-img0006.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - p62/SQSTM1 Antibody [NBP1-48320] - Immunoblotting analysis of LC3 and p62/SQSTM1 expression in thioglycollate-elicited peritoneal macrophages. Macrophages were isolated from C57BL/6 mice fed diets for 12 weeks, treated with or without 100 ng/ml LPS for 18 h with or without 50 nM bafilomycin A1. Image collected and cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S0022227520339523), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Western-Blot-NBP1-48320-img0012.jpg)
![<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - p62/SQSTM1 Antibody [NBP1-48320] - Untreated HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-p628/SQSTM1 at a 1:200 dilution overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Immunocytochemistry-Immunofluorescence-NBP1-48320-img0009.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - p62/SQSTM1 Antibody [NBP1-48320] - Vacuolin-1 analogues identified via virtual screening inhibited autophagic flux in HeLa cells. (A) Treatment of HeLa cells with vacuolin-1 analogues (10 M) and BAF (100 nM) failed to further increase the accumulation of both LC3B-II and SQSTM1 as compared to either drug alone. (B) Vacuolin-1 (10 M) or VS6 (10 M) induced the accumulation of yellow LC3B-II puncta in RFP-GFP-LC3B expressing HeLa cells. Scale bar = 10 m. Identification of Novel Vacuolin-1 Analogues as Autophagy Inhibitors by Virtual Drug Screening and Chemical Synthesis. Molecules (2017)](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Western-Blot-NBP1-48320-img0010.jpg)
![Western Blot: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - p62/SQSTM1 Antibody [NBP1-48320] - Vacuolin-1 analogues identified via virtual screening induced the accumulation of both LC3B-II and SQSTM1 in HeLa cells in a dose dependent manner after a 6 h treatment. Identification of Novel Vacuolin-1 Analogues as Autophagy Inhibitors by Virtual Drug Screening and Chemical Synthesis. Molecules (2017)](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Western-Blot-NBP1-48320-img0011.jpg)
![Simple Western: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - Drp1+/- mice is protective against autophagy impairment induced by Mn. (A) Representative images of the coronal mouse midbrain section (20um) co-immunostained for DA neurons (TH, red) in the SNpc and GABA neurons (GAD67, green) in the SNpr. Both of these brain regions were removed by laser microdissection for immunoblotting of p62 (top panels) in DA neurons (B) and GABA neurons (C). Total proteins per lane (bottom panels) were used as loading control. (D) Quantified levels of p62 were significantly increased in DA neurons (P = 0.0013), but not in the GABA neurons (P = 0.5457), of the Mn-treated WT mice. Mn did not significantly increase p62 in DA neurons of the Drp1+/- mice (P = 0.8660) No significant baseline level differences between the two genotypes were observed (P = 0.5664 for TH neurons and P = 0.7675 for GABA neurons. Data represent mean +- SEM, n = 5 for WT control, n = 6 for other groups), two-way ANOVA followed by Tukey post-hoc test. (E) Representative confocal images of mitochondrial morphology of DA neurons after TOM20 immunostaining (upper panels), then skeletonized (middle panels) for subsequent analysis. Scale bar 20um. (F, G) Various parameters of mitochondrial morphology were quantified using Fiji MiNA plugin. Data represents mean +- SEM (n = 6 mice per group), analyzed by one-way ANOVA, followed by Tukey post-hoc test Image collected and cropped by CiteAb from the following publication (//molecularneurodegeneration.biomedcentral.com/articles/10.1186/s13024-024-00708-w ), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-48320_rabbit-polyclonal-p62-sqstm1-antibody-308202410445514.jpg)
![Simple Western: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - Drp1+/- mice is protective against autophagy impairment induced by Mn. (A) Representative images of the coronal mouse midbrain section (20um) co-immunostained for DA neurons (TH, red) in the SNpc and GABA neurons (GAD67, green) in the SNpr. Both of these brain regions were removed by laser microdissection for immunoblotting of p62 (top panels) in DA neurons (B) and GABA neurons (C). Total proteins per lane (bottom panels) were used as loading control. (D) Quantified levels of p62 were significantly increased in DA neurons (P = 0.0013), but not in the GABA neurons (P = 0.5457), of the Mn-treated WT mice. Mn did not significantly increase p62 in DA neurons of the Drp1+/- mice (P = 0.8660) No significant baseline level differences between the two genotypes were observed (P = 0.5664 for TH neurons and P = 0.7675 for GABA neurons. Data represent mean +- SEM, n = 5 for WT control, n = 6 for other groups), two-way ANOVA followed by Tukey post-hoc test. (E) Representative confocal images of mitochondrial morphology of DA neurons after TOM20 immunostaining (upper panels), then skeletonized (middle panels) for subsequent analysis. Scale bar 20um. (F, G) Various parameters of mitochondrial morphology were quantified using Fiji MiNA plugin. Data represents mean +- SEM (n = 6 mice per group), analyzed by one-way ANOVA, followed by Tukey post-hoc test Image collected and cropped by CiteAb from the following publication (//molecularneurodegeneration.biomedcentral.com/articles/10.1186/s13024-024-00708-w ), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-48320_rabbit-polyclonal-p62-sqstm1-antibody-30820241044554.jpg)
![Immunocytochemistry/ Immunofluorescence: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - Effects on autophagic activity & dendritic length of neurons exposed to supernatant from HIV-1-infected microglia in combination with morphine. (A) Representative images of neurons with the indicated treatments. Sup, supernatant from uninfected [HIV(-)] & HIV-1-infected [HIV(+)] microglia. Cells were immunolabeled with antibodies to the autophagic activity marker p62/SQSTM1 (red) & the neuronal cell-type-specific marker MAP2 (green). DAPI (blue) staining indicates cell nuclei. (B) Quantification of p62/SQSTM1 immunoreactivity from (A). Data are presented as the percentage of control cells which was set at 100; F(5,24) = 5.882, p = 0.0011; ∗p < 0.05 when compared to HIV(+)Sup + morphine treatment. (C) Western blotting analysis of p62/SQSTM1 & LAMP1 expression levels for the indicated treatments. GAPDH was used as a loading control. Blots are representative of three independent experiments. (D) Measurement of dendrite length from (A). F(5,24) = 26.15, p = < 0.0001; Φp < 0.05 when compared to morphine; Ψp < 0.05 when compared to HIV(-)Sup; Ωp < 0.05 when compared to HIV(-)Sup + morphine; #p < 0.05 when compared to HIV(+)Sup; & ∗p < 0.05 when compared to HIV(+)Sup + morphine treatment. Error bars show the SEM for five randomly selected fields totaling at least 100 cells from each group. Image collected & cropped by CiteAb from the following publication (//journal.frontiersin.org/Article/10.3389/fmicb.2015.00653/abstract), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-48320_rabbit-polyclonal-p62-sqstm1-antibody-310202416163716.jpg)
![Western Blot: p62/SQSTM1 Antibody [NBP1-49956] - p53 repression of autophagy is PINK1-dependent. LC3-2/LC3-1 (a,b, N = 12), p62/SQSTM1 (a,c, N = 12), TIM23 (a,d, N = 12), TOM20 (a,e, N = 12) and HSP60 (a,f, N = 12) protein levels were analyzed in control (PINK+/+) or Pink1-deficient (PINK-/-) HAP cells treated with vehicle (Ct), pifithrin-alpha (PFT, 30 uM, 4 h) or CCCP (CP, 10 uM, 6 h) as described in the Methods. Bars represent the means +/- SEM of 4 independent experiments performed in triplicates and are expressed as percent of control HAP1 (HAP+/+, Ct) cells. Actin expression is provided as a representative gel loading control. Statistical analyses were performed with GraphPad Prism software by using unpaired Student's t-test. Significant differences are: ns not significant, *p < 0.05, ** p < 0.01 and ***p < 0.001. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41418-017-0016-0), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Western-Blot-NBP1-49956-img0006.jpg "Western Blot: p62/SQSTM1 Antibody [NBP1-49956] - p53 repression of autophagy is PINK1-dependent. LC3-2/LC3-1 (a,b, N = 12), p62/SQSTM1 (a,c, N = 12), TIM23 (a,d, N = 12), TOM20 (a,e, N = 12) and HSP60 (a,f, N = 12) protein levels were analyzed in control (PINK+/+) or Pink1-deficient (PINK-/-) HAP cells treated with vehicle (Ct), pifithrin-alpha (PFT, 30 uM, 4 h) or CCCP (CP, 10 uM, 6 h) as described in the Methods. Bars represent the means +/- SEM of 4 independent experiments performed in triplicates and are expressed as percent of control HAP1 (HAP+/+, Ct) cells. Actin expression is provided as a representative gel loading control. Statistical analyses were performed with GraphPad Prism software by using unpaired Student's t-test. Significant differences are: ns not significant, *p < 0.05, ** p < 0.01 and ***p < 0.001. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41418-017-0016-0), licensed under a CC-BY license.")
![Immunohistochemistry-Paraffin: p62/SQSTM1 Antibody [NBP1-49956] - Section of human breast carcinoma. Antibody: Affinity purified rabbit anti- Sequestosome-1 used at a dilution of 1:200 (1ug/ml). Detection: Vector Laboratories ImmPACT NovaRED Peroxidase Substrate](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Immunohistochemistry-Paraffin-NBP1-49956-img0004.jpg)
![<b>Independent Antibodies Validation and Biological Strategies Validation.</b>Western Blot: p62/SQSTM1 Antibody [NBP1-49956] - Detection of Human and Mouse Sequestosome-1 by Western Blot (h&m) and Immunoprecipitation (h). Samples: Whole cell lysate from HeLa (5, 15 and 50 mcg for WB; 1 mg for IP, 20% of IP loaded), 293T (T; 50 mcg), and mouse NIH3T3 (M; 50 mcg) cells. Antibodies: Affinity purified rabbit anti-Sequestosome-1 antibody used for WB at 0.04 mcg/ml (A) and 0.4 mcg/ml (B) and used for IP at 3 mcg/mg lysate. Sequestosome-1 was also immunoprecipitated by rabbit anti-Sequestosome-1 antibodies which recognize upstream epitopes. For blotting immunoprecipitated Sequestosome-1 was used. Detection: Chemiluminescence with exposure times of 30 seconds (A) and 3 seconds (B).](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Western-Blot-NBP1-49956-img0003.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: p62/SQSTM1 Antibody [NBP1-49956] - Evaluation of mitophagy control by p53 in mice brain. p62/SQSTM1 (N = 9) protein levels were analyzed in control (p53+/+) or TP53-deficient (p53-/-) knockout mice brains as described in the Methods. Bars represent the means +/- SEM of 3 independent experiments performed in triplicate and are expressed as percent of control (p53+/+) cells. Actin expression is provided as a representative gel loading control. Statistical analyses were performed with GraphPad Prism software by using unpaired Student's t-test. Significant differences are: ** p < 0.01, ***p < 0.001. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41418-017-0016-0), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Western-Blot-NBP1-49956-img0005.jpg)
![Western Blot: p62/SQSTM1 Antibody [NBP1-42821] - HeLa cells were treated with (+) or without 50 uM (-) of Chloriquine (CQ) for 24 hours. Total cell lysates were prepared and separated on a 12% gel by SDS-PAGE. Protein was transferred to PVDF membrane and blocked in 5% non-fat milk. The membrane was then probed with 2 ug/ml anti-p62/SQSMT1 in 1% milk and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. Note the upregulation of p62 (arrowhead) in response to chloroquine treatment and the blockage of autophagy.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Western-Blot-NBP1-42821-img0007.jpg "<b>Biological Strategies Validation. </b>Western Blot: p62/SQSTM1 Antibody [NBP1-42821] - HeLa cells were treated with (+) or without 50 uM (-) of Chloriquine (CQ) for 24 hours. Total cell lysates were prepared and separated on a 12% gel by SDS-PAGE. Protein was transferred to PVDF membrane and blocked in 5% non-fat milk. The membrane was then probed with 2 ug/ml anti-p62/SQSMT1 in 1% milk and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. Note the upregulation of p62 (arrowhead) in response to chloroquine treatment and the blockage of autophagy.")
![<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody [NBP1-42821] - Confocal immunofluorescent analysis of HeLa cells using p62/SQSTM1 antibody (NBP1-42821, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Immunocytochemistry-Immunofluorescence-NBP1-42821-img0006.jpg)
![Western Blot: p62/SQSTM1 Antibody [NBP1-42821] - Western Blot of the lysate from primary rat hepatocytes. Image from verified customer review.](http://images.novusbio.com/fullsize/antibody/nbp1-42821_rabbit-polyclonal-p62-sqstm1-antibody-western-blot-672023145728..jpg)
![Western Blot: p62/SQSTM1 Antibody [NBP1-42821] - Western Blot of the lysate from mice hepatocytes. Image from verified customer review.](http://images.novusbio.com/fullsize/antibody/nbp1-42821_rabbit-polyclonal-p62-sqstm1-antibody-western-blot-672023145955..jpg)
![Western Blot: p62/SQSTM1 Antibody [NBP1-42821] - Western Blot of the lysate of HepG2 cells. Image from verified customer review.](http://images.novusbio.com/fullsize/antibody/nbp1-42821_rabbit-polyclonal-p62-sqstm1-antibody-western-blot-67202315152..jpg)
![Knockout Validated: p62/SQSTM1 Antibody [NBP1-42822] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and p62/SQSTM1 knockout (KO) HeLa cell line. PVDF membrane was probed with Rabbit Anti-Human p62/SQSTM1 polyclonal Antibody (Catalog # NBP1-42822) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog #HAF008). Specific band was detected for p62/SQSTM1 at approximately 60 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. This experiment was conducted under reducing conditions.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Knockout-Validated-NBP1-42822-img0009.jpg "<b>Genetic Strategies Validation. </b>Knockout Validated: p62/SQSTM1 Antibody [NBP1-42822] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and p62/SQSTM1 knockout (KO) HeLa cell line. PVDF membrane was probed with Rabbit Anti-Human p62/SQSTM1 polyclonal Antibody (Catalog # NBP1-42822) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog #HAF008). Specific band was detected for p62/SQSTM1 at approximately 60 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. This experiment was conducted under reducing conditions.")
![<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Western Blot: p62/SQSTM1 Antibody [NBP1-42822] - HeLa cells were treated with (+) or without 50 uM (-) of Chloriquine (CQ) for 24 hours. Total cell lysates were prepared and separated on a 12% gel by SDS-PAGE. Protein was transferred to PVDF membrane and blocked in 5% non-fat milk. The membrane was then probed with 2 ug/ml anti-p62/SQSMT1 in 1% milk and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. Note the upregulation of p62 (arrowhead) in response to chloroquine treatment and the blockage of autophagy.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Western-Blot-NBP1-42822-img0008.jpg)
![Simple Western: p62/SQSTM1 Antibody [NBP1-42822] - Simple Western lane view shows a specific band for p62/SQSTM1 in 0.5 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Simple-Western-NBP1-42822-img0007.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: p62/SQSTM1 Antibody [NBP1-42822] - Hyperosmotic stress stimulatesA utophagyHeLa and HCT 116 cells were transduced with an adenovirus coding for GFP-LC3 (Ad GFP-LC3) for 24 h. LC3 I-to-LC3 II conversion and p62/SQSTM1 depletion were assessed by Western blot analysis in HeLa and HCT116 cells treated with various concentrations of sorbitol or mannitol (25-300 mOsm) .It indicated times (0.5-6 h). Image collected and cropped by Citeab from the following publication ( licensed under a CC-BY license.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Western-Blot-NBP1-42822-img0010.jpg)
![Western Blot: p62/SQSTM1 Antibody [NBP1-49954] - Human AC16 under normoxic and hypoxic conditions. WB image submitted by a verified customer review.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Western-Blot-NBP1-49954-img0006.jpg "<b>Biological Strategies Validation. </b>Western Blot: p62/SQSTM1 Antibody [NBP1-49954] - Human AC16 under normoxic and hypoxic conditions. WB image submitted by a verified customer review.")
![<b>Biological Strategies Validation. </b>Immunohistochemistry-Paraffin: p62/SQSTM1 Antibody [NBP1-49954] - Sample: FFPE section of human breast carcinoma. Antibody: Affinity purified rabbit anti- Sequestosome-1 used at a dilution of 1:1,000 (1ug/ml). Detection: Vector Laboratories ImmPACT NovaRED Peroxidase Substrate](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Immunohistochemistry-NBP1-49954-img0005.jpg)
![<b>Independent Antibodies Validation and Biological Strategies Validation.</b>Western Blot: p62/SQSTM1 Antibody [NBP1-49954] - Detection of Human and Mouse Sequestosome-1 by Western Blot (h&m) and Immunoprecipitation (h). Samples: Whole cell lysate from HeLa (5, 15 and 50 mcg for WB; 1 mg for IP, 20% of IP loaded), 293T (T; 50 mcg), and mouse NIH3T3 (M; 50 mcg) cells. Antibodies: Affinity purified rabbit anti-Sequestosome-1 antibody used for WB at 0.1 mcg/ml (A) and 0.4 mcg/ml (B) and used for IP at 3 mcg/mg lysate. Sequestosome-1 was also immunoprecipitated by rabbit anti-Sequestosome-1 antibodies which recognize downstream epitopes. For blotting immunoprecipitated Sequestosome-1 was used. Detection: Chemiluminescence with exposure times of 30 seconds (A) and 3 seconds (B).](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Western-Blot-NBP1-49954-img0004.jpg)
![Simple Western: p62/SQSTM1 Antibody [NBP1-49954] - Simple Western lane view shows a specific band for p62/SQSTM1 in 0.5 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Simple-Western-NBP1-49954-img0003.jpg)
![Knockout Validated: p62/SQSTM1 Antibody (2533B) [NBP2-92995] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and p62/SQSTM1 knockout HeLa cell line (KO). PVDF membrane was probed with 1 ug/mL of Rabbit Anti-Human p62/SQSTM1 Monoclonal Antibody (Catalog # NBP2-92995) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for p62/SQSTM1 at approximately 60 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # 2275-PC-100) is shown as a loading control. This experiment was conducted under reducing conditions.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-2533B-Knockout-Validated-NBP2-92995-img0006.jpg "<b>Genetic Strategies Validation. </b>Knockout Validated: p62/SQSTM1 Antibody (2533B) [NBP2-92995] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and p62/SQSTM1 knockout HeLa cell line (KO). PVDF membrane was probed with 1 ug/mL of Rabbit Anti-Human p62/SQSTM1 Monoclonal Antibody (Catalog # NBP2-92995) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for p62/SQSTM1 at approximately 60 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # 2275-PC-100) is shown as a loading control. This experiment was conducted under reducing conditions.")
![<b>Genetic Strategies Validation. </b>Western Blot: p62/SQSTM1 Antibody (2533B) [NBP2-92995] - Western blot shows lysates of A172 human glioblastoma cell line and RT-4 human bladder carcinoma cell line. PVDF membrane was probed with 1 ug/mL of Rabbit Anti-Human p62/SQSTM1 Monoclonal Antibody (Catalog # NBP2-92995) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for p62/SQSTM1 at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-2533B-Western-Blot-NBP2-92995-img0004.jpg)
![Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody (2533B) [NBP2-92995] - p62/SQSTM1 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with chloroquine (left) or untreated (right) using Rabbit Anti-Human p62/SQSTM1 Monoclonal Antibody (Catalog # NBP2-92995) at 8 ug/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-2533B-Immunocytochemistry-Immunofluorescence-NBP2-92995-img0002.jpg)
![Immunohistochemistry-Paraffin: p62/SQSTM1 Antibody (2533B) [NBP2-92995] - p62/SQSTM1 was detected in immersion fixed paraffin-embedded sections of human liver using Rabbit Anti-Human p62/SQSTM1 Monoclonal Antibody (Catalog #NBP2-92995) at 10 ug/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell nuclei.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-2533B-Immunohistochemistry-Paraffin-NBP2-92995-img0003.jpg)
![Simple Western: p62/SQSTM1 Antibody (2533B) [NBP2-92995] - Simple Western lane view shows lysates of RT-4 human bladder carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for p62/SQSTM1 at approximately 64 kDa (as indicated) using 20 ug/mL of Rabbit Anti-Human p62/SQSTM1 Monoclonal Antibody (Catalog # NBP2-92995). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-2533B-Simple-Western-NBP2-92995-img0005.jpg)
![ELISA: p62/SQSTM1 Antibody (1C9) [H00008878-M03] - Detection limit for recombinant GST tagged SQSTM1 is approximately 3ng/ml as a capture antibody.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-1C9-Sandwich-ELISA-H00008878-M03-img0002.jpg "ELISA: p62/SQSTM1 Antibody (1C9) [H00008878-M03] - Detection limit for recombinant GST tagged SQSTM1 is approximately 3ng/ml as a capture antibody.")
![Western Blot: p62/SQSTM1 Antibody (1478) [NBP2-43663] - Non-transfected (-) and transfected (+) HepG2 whole cell extracts (50 ug) were separated by 10% SDS-PAGE, and the membrane was blotted with SQSTM1 antibody (1478) dilution 1:500.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-1478-Western-Blot-NBP2-43663-img0008.jpg "<b>Genetic Strategies Validation. </b>Western Blot: p62/SQSTM1 Antibody (1478) [NBP2-43663] - Non-transfected (-) and transfected (+) HepG2 whole cell extracts (50 ug) were separated by 10% SDS-PAGE, and the membrane was blotted with SQSTM1 antibody (1478) dilution 1:500.")
![<b>Genetic Strategies Validation. </b>Immunohistochemistry-Frozen: p62/SQSTM1 Antibody (1478) [NBP2-43663] - Frozen sectioned adult mouse retina. Green: SQSTM1 protein stained by SQSTM1 antibody [1478] diluted at 1:250. Red: beta Tubulin 3/ TUJ1, stained by beta Tubulin 3/ TUJ1 antibody [11710] (NBP2-43559) diluted at 1:250. Blue: Fluoroshield with DAPI.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-1478-Immunohistochemistry-Frozen-NBP2-43663-img0014.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: p62/SQSTM1 Antibody (1478) [NBP2-43663] - Analysis of un-treated (-) and treated (+), Thapsigargin treatment for 12hr and 24hr. HepG2 whole cell extracts (30 ug) were separated by 10% SDS-PAGE, and the membrane was blotted with SQSTM1 antibody (1478) dilution 1:1000. Beta Actin was used as internal control (bottom panel).](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-1478-Western-Blot-NBP2-43663-img0002.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: p62/SQSTM1 Antibody (1478) [NBP2-43663] - Analysis of un-treated (-) and treated (+), Thapsigargin treatment for 12hr and 24hr. Huh-7 whole cell extracts (30 ug) were separated by 10% SDS-PAGE, and the membrane was blotted with SQSTM1 antibody (1478) dilution 1:1000. Beta Actin was used as internal control (bottom panel).](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-1478-Western-Blot-NBP2-43663-img0003.jpg)
![<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody (1478) [NBP2-43663] - Analysis of Samples: HeLa cells mock (left) and treated with 50uM Chloroquine for 24 hr (right) were fixed in 4% paraformaldehyde at RT for 15 min. Green: SQSTM1 protein stained by SQSTM1 antibody [1478]. Red: phalloidin, a cytoskeleton marker, stained by phalloidin diluted at 1:200. Blue: Hoechst 33342 staining.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-1478-Immunocytochemistry-Immunofluorescence-NBP2-43663-img0006.jpg)
![Flow Cytometry: p62/SQSTM1 Antibody (1478) [NBP2-43663] - Analysis of HeLa cell. Brown: Unlabelled sample was also used as a control. Blue: SQSTM1 antibody [1478] dilution: 1:100.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-1478-Flow-Cytometry-NBP2-43663-img0004.jpg)
![Western Blot: p62/SQSTM1 Antibody [NBP1-49955] - Detection of Human Sequestosome-1 by Western Blot and Immunoprecipitation. Samples: Whole cell lysate (5, 15 and 50 mcg for WB; 1 mg for IP, 20% of IP loaded) from HeLa cells. Antibodies: Affinity purified rabbit anti-Sequestosome-1 antibody used for WB at 0.1 mcg/ml (A) and 0.4 mcg/ml (B) and used for IP at 3 mcg/mg lysate. Sequestosome-1 was also immunoprecipitated by rabbit anti-Sequestosome-1 antibodies which recognize other epitopes. For blotting immunoprecipitated Sequestosome-1 was used. Detection: Chemiluminescence with exposure times of 30 seconds (A) and 3 seconds (B).](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Western-Blot-NBP1-49955-img0002.jpg "<b>Independent Antibodies Validation. </b>Western Blot: p62/SQSTM1 Antibody [NBP1-49955] - Detection of Human Sequestosome-1 by Western Blot and Immunoprecipitation. Samples: Whole cell lysate (5, 15 and 50 mcg for WB; 1 mg for IP, 20% of IP loaded) from HeLa cells. Antibodies: Affinity purified rabbit anti-Sequestosome-1 antibody used for WB at 0.1 mcg/ml (A) and 0.4 mcg/ml (B) and used for IP at 3 mcg/mg lysate. Sequestosome-1 was also immunoprecipitated by rabbit anti-Sequestosome-1 antibodies which recognize other epitopes. For blotting immunoprecipitated Sequestosome-1 was used. Detection: Chemiluminescence with exposure times of 30 seconds (A) and 3 seconds (B).")
![<b>Independent Antibodies Validation. </b>Immunohistochemistry-Paraffin: p62/SQSTM1 Antibody [NBP1-49955] - Section of human lung carcinoma. Antibody: Affinity purified rabbit anti- Sequestosome-1 used at a dilution of 1:1,000 (1ug/ml). Detection: Vector Laboratories ImmPACT NovaRED Peroxidase Substrate](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Immunohistochemistry-NBP1-49955-img0003.jpg)
![Western Blot: p62/SQSTM1 Antibody (PS00-61) [NBP3-32702] - Western blot analysis of p62/SQSTM1 on different lysates with Rabbit anti-p62/SQSTM1 antibody (NBP3-32702) at 1/500 dilution. Lane 1: PC-12 cell lysate (10 ug/Lane) Lane 2: RAW264.7 cell lysate (10 ug/Lane) Lane 3: PANC-1 cell lysate (10 ug/Lane) Lane 4: Mouse liver tissue lysate (20 ug/Lane) Lane 5: Rat brain tissue lysate (20 ug/Lane) Predicted band size: 48 kDa Observed band size: 62 kDa Exposure time: 2 minutes; 8% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32702) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:300,000 dilution was used for 1 hour at room temperature.](http://images.novusbio.com/fullsize/nbp3-32702_rabbit-p62-sqstm1-mab-ps00-61-278202415295572.jpg "Western Blot: p62/SQSTM1 Antibody (PS00-61) [NBP3-32702] - Western blot analysis of p62/SQSTM1 on different lysates with Rabbit anti-p62/SQSTM1 antibody (NBP3-32702) at 1/500 dilution. Lane 1: PC-12 cell lysate (10 ug/Lane) Lane 2: RAW264.7 cell lysate (10 ug/Lane) Lane 3: PANC-1 cell lysate (10 ug/Lane) Lane 4: Mouse liver tissue lysate (20 ug/Lane) Lane 5: Rat brain tissue lysate (20 ug/Lane) Predicted band size: 48 kDa Observed band size: 62 kDa Exposure time: 2 minutes; 8% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32702) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:300,000 dilution was used for 1 hour at room temperature.")
![Immunohistochemistry: p62/SQSTM1 Antibody (PS00-61) [NBP3-32702] - Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-p62/SQSTM1 antibody (NBP3-32702) at 1/400 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32702) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.](http://images.novusbio.com/fullsize/nbp3-32702_rabbit-p62-sqstm1-mab-ps00-61-298202414161843.jpg)
![Immunocytochemistry/ Immunofluorescence: p62/SQSTM1 Antibody (PS00-61) [NBP3-32702] - Immunocytochemistry analysis of A549 cells labeling p62/SQSTM1 with Rabbit anti-p62/SQSTM1 antibody (NBP3-32702) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-p62/SQSTM1 antibody (NBP3-32702) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.](http://images.novusbio.com/fullsize/nbp3-32702_rabbit-p62-sqstm1-mab-ps00-61-29202410484897.jpg)
![Western Blot: p62/SQSTM1 Antibody [NBP2-56317] - Western blot analysis in human cell line RT-4, human cell line U-251 MG, human plasma and human liver tissue.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Western-Blot-NBP2-56317-img0002.jpg "Western Blot: p62/SQSTM1 Antibody [NBP2-56317] - Western blot analysis in human cell line RT-4, human cell line U-251 MG, human plasma and human liver tissue.")
![Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody [NBP2-56317] - Staining of human cell line A549 shows localization to cytosol & vesicles.](http://images.novusbio.com/fullsize/p62-SQSTM1-Antibody-Immunocytochemistry-Immunofluorescence-NBP2-56317-img0001.jpg)