SQSTM1 / p62 antibody | knockout validation | Abcam ab56416

This is a knockout-validated antibody summary, based on publications "UXT chaperone prevents proteotoxicity by acting as an autophagy adaptor for p62-dependent aggrephagy", as cited below [1], and "MOAP-1-mediated dissociation of p62/SQSTM1 bodies releases Keap1 and suppresses Nrf2 signaling" for (figure 1e, 2c,1f, 2i) [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

SQSTM1 / p62 antibody | knockout validation | Abcam ab56416 figure 1
Figure 1. p62 knockout HeLa cells (HeLa/p62KO) were transiently transfected with FLAG-tagged p62 (wild type or ΔZZ–LB). Cells were treated with 5µM MG132 for 6h, and stained with anti-ubiquitin and anti-p62 antibodies followed by anti-rabbit IgG-TRITC and anti-mouse IgG-Alexa-350 antibodies. DIC differential interference contrast images. Scale bar, 10µm. From [1].
Antibody information

Mouse monoclonal IgG2a

Company: Abcam

Antibody: SQSTM1 / p62

Catalog number: ab56416

Summary: Mouse monoclonal antibody against recombinant full length protein corresponding to human SQSTM1/ p62 (aa 1-440). Reacts with human. Suitable for western blot, immunoprecipitation, immunocytochemistry/immunofluorescence, immunohistochemistry (paraffin) and flow cytometry.

Validation Method

Immunocytochemistry

Sample

Control and p62 knockout HeLa cells.

Blocking agent

1% BSA, 22.52mg/mL glycine, and 0.1% gelatin in PBST

Primary incubation

1:500 dilution.

Secondary incubation

1:1,000 dilution anti-mouse IgG-Alexa Fluor 350 (Thermo Fisher, A11068) in blocking solution.

Detection

Images were acquired using a fluorescence microscope (Ti-E, Nikon) equipped with a 100× (1.4N.A) Plan-Apochromat objective lens and a charge-coupled camera device (DS-Qi2, Nikon).

References
  1. Yoon M, Choi B, Kim E, Ohk J, Yang C, Choi Y, et al. UXT chaperone prevents proteotoxicity by acting as an autophagy adaptor for p62-dependent aggrephagy. Nat Commun. 2021;12:1955 pubmed publisher
  2. Tan C, Chang H, Zhou Q, Yu C, Fu N, Sabapathy K, et al. MOAP-1-mediated dissociation of p62/SQSTM1 bodies releases Keap1 and suppresses Nrf2 signaling. EMBO Rep. 2021;:e50854 pubmed publisher