Invitrogen

SQSTM1 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated. View Details

9 Published Figures

11 References

View all (74) SQSTM1 antibodies

Datasheet

Protocols

Questions & Answers

Tech Support

Datasheet

Protocols

Questions & Answers

Tech Support

Product Image Gallery

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) — Western blot was performed using Anti-SQSTM1 Polyclonal Antibody (Product # PA5-20839) and a ~55 kDa band corresponding to SQSTM1 was observed across cell lines and tissue tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1),Hep G2 (Lane 2), BeWo (Lane 3), SH-SY5Y (Lane 4) and THP 1 (Lane 5); and tissue extracts of Mouse Thymus were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (0.75 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

SQSTM1 Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. Hep G2 cells were transfected with SQSTM1 siRNA and decrease in signal intensity was observed in Western Blot application using Anti-SQSTM1 Polyclonal Antibody (Product # PA5-20839). {KD}

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

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

0.5-2 µg/mL

View 7 publications 7 publications

Immunohistochemistry (IHC)

2-5 µg/mL

View 1 publication 1 publication

Immunocytochemistry (ICC/IF)

20 µg/mL

View 3 publications 3 publications

Product Specifications

Species Reactivity

Human, Mouse, Rat

Published species

Human, Mouse

Host/Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

A 13 amino acid peptide from near the carboxy terminus of human SQSTM1.

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Antigen affinity chromatography

Storage buffer

PBS

Contains

0.02% sodium azide

Storage conditions

Maintain refrigerated at 2-8°C for up to 3 months. For long term storage store at -20°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.

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)