Invitrogen

SQSTM1 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated.

13 Published Figures

14 References

View all (82) SQSTM1 antibodies

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Datasheet

Protocols

Questions & Answers

Tech Support

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SQSTM1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of SQSTM1 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with SQSTM1 Polyclonal Antibody (Product # PA5-20839) at 20 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cytoplasmic, Endosome-like, plasma membrane and nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

SQSTM1 Antibody in Immunohistochemistry (IHC) — Immunohistochemical analysis of paraffin-embedded Human Spleen Tissue using SQSTM1 Polyclonal Antibody (Product # PA5-20839) at 5 µg/mL. Tissue was fixed with formaldehyde and blocked with 0.1 serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

SQSTM1 Antibody in Western Blot (WB) — Knockout of SQSTM1 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR970861_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of SQSTM1 was performed by loading 30 µg of HeLa Wild Type (Lane 1), HeLa Cas9 (Lane 2) andHela SQSTM1 KO (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-SQSTM1 Polyclonal Antibody (Product # PA5-20839, 0.75 µg/mL dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:6,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to SQSTM1. Uncharacterized band was observed in all the samples at 60 kDa.

SQSTM1 Antibody in Western Blot, Immunocytochemistry (WB, ICC/IF) — delta-TT triggers autophagy in PC3, but not in DU145, prostate cancer cells. DU145 and PC3 cells were treated with delta-TT (15 mug/mL) for 1-24 h. A, Western blot analysis was performed to investigate the expression levels of autophagy-related proteins (LC3-II/LC3-I, SQSTM1/p62). Tubulin expression was evaluated as a loading control. B, The expression levels and intracellular localization of LC3 and SQSTM1/p62 were evaluated by immunofluorescence. One representative of three different experiments performed is shown. C, controls (vehicle). Scale bars are 20 mum

SQSTM1 Antibody — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. A loss of signal was observed for target protein inSQSTM1 KO cell line compared to control cell line using Anti-SQSTM1 Polyclonal Antibody (Product # PA5-20839). {KO}

Product Details

PA5-20839

Applications	Tested Dilution	Publications
Western Blot (WB)	0.5-2 µg/mL	View 8 publications 8 publications
Immunohistochemistry (IHC)	2-5 µg/mL	View 1 publication 1 publication
Immunocytochemistry (ICC/IF)	20 µg/mL	View 5 publications 5 publications

Product Specifications
Species Reactivity	Human, Mouse, Rat
Published species	Human, Mouse, Rodent
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	A 13 amino acid peptide from near the carboxy terminus of human SQSTM1. View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS
Contains	0.02% sodium azide
Storage conditions	4° C
RRID	AB_11157045

Product Specific Information

A suggested positive control is human spleen tissue lysate.

PA5-20839 can be used with blocking peptide PEP-0953.

This antibody has been tested by Western Blot in human and mouse samples; immunofluorescence and immunohistochemistry in human, mouse and rat samples. All other applications and species not yet tested.

Target Information

This gene encodes a multifunctional protein that binds ubiquitin and regulates activation of the nuclear factor kappa-B signaling pathway. The protein functions as a scaffolding/adaptor protein in concert with TNF receptor-associated factor 6 to mediate activation of NF-kB in response to upstream signals. Alternatively spliced transcript variants encoding either the same or different isoforms have been identified for this gene. Mutations in this gene result in sporadic and familial Paget disease of bone.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

NBR1 is a critical step in the repression of thermogenesis of p62-deficient adipocytes through PPARγ.

Nature communications

Huang J,Linares JF,Duran A,Xia W,Saltiel AR,Müller TD,Diaz-Meco MT,Moscat J

Published figure using SQSTM1 polyclonal antibody (Product # PA5-20839) in Western Blot

Mon May 17 00:00:00 EDT 2021

NBR1 is a critical step in the repression of thermogenesis of p62-deficient adipocytes through PPARγ.

Nature communications

Huang J,Linares JF,Duran A,Xia W,Saltiel AR,Müller TD,Diaz-Meco MT,Moscat J

Published figure using SQSTM1 polyclonal antibody (Product # PA5-20839) in Western Blot

Mon May 17 00:00:00 EDT 2021

Chick Embryo: A Preclinical Model for Understanding Ischemia-Reperfusion Mechanism.

Frontiers in pharmacology

Fauzia E,Barbhuyan TK,Shrivastava AK,Kumar M,Garg P,Khan MA,Robertson AAB,Raza SS

Published figure using SQSTM1 polyclonal antibody (Product # PA5-20839) in Western Blot

Thu Oct 01 00:00:00 EDT 2020

Sinoporphyrin sodium is a promising sensitizer for photodynamic and sonodynamic therapy in glioma.

Oncology reports

An YW,Liu HQ,Zhou ZQ,Wang JC,Jiang GY,Li ZW,Wang F,Jin HT

PA5-20839 was used in Western Blotting to explore the antitumor effects of sinoporphyrin sodium (DVDMS)‚mediated photodynamic therapy (PDT) and sonodynamic therapy (SDT) in glioma, and to reveal the underlying mechanisms.

Thu Oct 01 00:00:00 EDT 2020

The Second Case of Saposin A Deficiency and Altered Autophagy.

JIMD reports

Kose M,Demir SA,Akinci G,Eraslan C,Yilmaz U,Ceylaner S,Yildirim ES,Seyrantepe V

PA5-20839 was used in Immunocytochemistry-immunoflourescence to report a case of Saposin A deficiency determined by a previously undescried homozygous variant in the saposin A domain of the PSAP gene.

Mon Sep 28 00:00:00 EDT 2020

Aggresome-Like Formation Promotes Resistance to Proteotoxicity in Cells from Long-Lived Species.

The journals of gerontology. Series A, Biological sciences and medical sciences

Sunchu B,Riordan RT,Yu Z,Almog I,Dimas-Munoz J,Drake AC,Perez VI

PA5-20839 was used in Immunocytochemistry to suggest that the resistance to proteotoxicity observed in long-lived species is not due to a lower level of protein aggregates but rather to enhanced handling of the protein aggregates through the formation of aggresome-like inclusions, a well-recognized protective mechanism against proteotoxicty.

Mon Jul 13 00:00:00 EDT 2020

Long Term Pharmacological Perturbation of Autophagy in Mice: Are HCQ Injections a Relevant Choice?

Biomedicines

Masson JD,Blanchet B,Periou B,Authier FJ,Mograbi B,Gherardi RK,Crépeaux G

Published figure using SQSTM1 polyclonal antibody (Product # PA5-20839) in Western Blot

Sun Mar 01 00:00:00 EST 2020

Long Term Pharmacological Perturbation of Autophagy in Mice: Are HCQ Injections a Relevant Choice?

Biomedicines

Masson JD,Blanchet B,Periou B,Authier FJ,Mograbi B,Gherardi RK,Crépeaux G

Published figure using SQSTM1 polyclonal antibody (Product # PA5-20839) in Western Blot

Sun Mar 01 00:00:00 EST 2020

δ-Tocotrienol induces apoptosis, involving endoplasmic reticulum stress and autophagy, and paraptosis in prostate cancer cells.

Cell proliferation

Fontana F,Moretti RM,Raimondi M,Marzagalli M,Beretta G,Procacci P,Sartori P,Montagnani Marelli M,Limonta P

Published figure using SQSTM1 polyclonal antibody (Product # PA5-20839) in Immunocytochemistry

Wed May 01 00:00:00 EDT 2019

δ-Tocotrienol induces apoptosis, involving endoplasmic reticulum stress and autophagy, and paraptosis in prostate cancer cells.

Cell proliferation

Fontana F,Moretti RM,Raimondi M,Marzagalli M,Beretta G,Procacci P,Sartori P,Montagnani Marelli M,Limonta P

Published figure using SQSTM1 polyclonal antibody (Product # PA5-20839) in Immunocytochemistry

Wed May 01 00:00:00 EDT 2019

δ-Tocotrienol induces apoptosis, involving endoplasmic reticulum stress and autophagy, and paraptosis in prostate cancer cells.

Cell proliferation

Fontana F,Moretti RM,Raimondi M,Marzagalli M,Beretta G,Procacci P,Sartori P,Montagnani Marelli M,Limonta P

Published figure using SQSTM1 polyclonal antibody (Product # PA5-20839) in Immunocytochemistry

Wed May 01 00:00:00 EDT 2019

δ-Tocotrienol induces apoptosis, involving endoplasmic reticulum stress and autophagy, and paraptosis in prostate cancer cells.

Cell proliferation

Fontana F,Moretti RM,Raimondi M,Marzagalli M,Beretta G,Procacci P,Sartori P,Montagnani Marelli M,Limonta P

Published figure using SQSTM1 polyclonal antibody (Product # PA5-20839) in Immunocytochemistry

Wed May 01 00:00:00 EDT 2019

Dual role of autophagy on docetaxel-sensitivity in prostate cancer cells.

Cell death & disease

Cristofani R,Montagnani Marelli M,Cicardi ME,Fontana F,Marzagalli M,Limonta P,Poletti A,Moretti RM

Published figure using SQSTM1 polyclonal antibody (Product # PA5-20839) in Western Blot

Thu Aug 30 00:00:00 EDT 2018

Dual role of autophagy on docetaxel-sensitivity in prostate cancer cells.

Cell death & disease

Cristofani R,Montagnani Marelli M,Cicardi ME,Fontana F,Marzagalli M,Limonta P,Poletti A,Moretti RM

Published figure using SQSTM1 polyclonal antibody (Product # PA5-20839) in Western Blot

Thu Aug 30 00:00:00 EDT 2018

Systemic overexpression of SQSTM1/p62 accelerates disease onset in a SOD1^H46R-expressing ALS mouse model.

Molecular brain

Mitsui S,Otomo A,Nozaki M,Ono S,Sato K,Shirakawa R,Adachi H,Aoki M,Sobue G,Shang HF,Hadano S

Published figure using SQSTM1 polyclonal antibody (Product # PA5-20839) in Immunohistochemistry

Tue May 29 00:00:00 EDT 2018

Systemic overexpression of SQSTM1/p62 accelerates disease onset in a SOD1^H46R-expressing ALS mouse model.

Molecular brain

Mitsui S,Otomo A,Nozaki M,Ono S,Sato K,Shirakawa R,Adachi H,Aoki M,Sobue G,Shang HF,Hadano S

Published figure using SQSTM1 polyclonal antibody (Product # PA5-20839) in Immunohistochemistry

Tue May 29 00:00:00 EDT 2018

Systemic overexpression of SQSTM1/p62 accelerates disease onset in a SOD1^H46R-expressing ALS mouse model.

Molecular brain

Mitsui S,Otomo A,Nozaki M,Ono S,Sato K,Shirakawa R,Adachi H,Aoki M,Sobue G,Shang HF,Hadano S

Published figure using SQSTM1 polyclonal antibody (Product # PA5-20839) in Immunohistochemistry

Tue May 29 00:00:00 EDT 2018

Ubiquitin-mediated regulation of the E3 ligase GP78 by MGRN1 in trans affects mitochondrial homeostasis.

Journal of cell science

Mukherjee R,Chakrabarti O

PA5-20839 was used in western blot to determine how mitochondrial homeostasis is affected by ubiquitin-mediated regulation of E3 ligase GP78 by MGRN1 in trans

Mon Feb 15 00:00:00 EST 2016

Mahogunin regulates fusion between amphisomes/MVBs and lysosomes via ubiquitination of TSG101.

Cell death & disease

Majumder P,Chakrabarti O

Published figure using SQSTM1 polyclonal antibody (Product # PA5-20839) in Western Blot

Thu Nov 05 00:00:00 EST 2015

Bioinformatics

Protein Aliases: EBI3-associated protein of 60 kDa; EBI3-associated protein p60; EBIAP; oxidative stress induced; oxidative stress induced like; phosphotyrosine independent ligand for the Lck SH2 domain p62; Phosphotyrosine-independent ligand for the Lck SH2 domain of 62 kDa; PKC-zeta-interacting protein; Protein kinase C-zeta-interacting protein; Sequestosome-1; STONE14; Ubiquitin-binding protein p62

Gene Aliases: A170; FTDALS3; ORCA; OSF-6; Osi; OSIL; p60; p62; p62B; PDB3; SQSTM1; STAP; STONE14; ZIP; ZIP3

UniProt ID: (Human) Q13501, (Rat) O08623, (Mouse) Q64337

Entrez Gene ID: (Human) 8878, (Rat) 113894, (Mouse) 18412

Function(s)