This is a knockout-validated antibody summary, based on the publication "p53-dependent autophagic degradation of TET2 modulates cancer therapeutic resistance", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Mouse monoclonal IgG1 (kappa light chain)

Company: Santa Cruz

Antibody: SQSTM1

Catalog number: sc-28359

Summary: Mouse monoclonal IgG1 (kappa light chain) against amino acids 151-440 of human SQSTM1 (p62). Recommended for detection of sequestosome 1 of human origin by western blot, immunoprecipitation, immunofluorescence, immunohistochemistry (paraffin) and ELISA.

Western blot

WT and p62KO p53-inducible HEK293T cells. Cells were lysed on ice by using RIPA lysis buffer (50mM Tris-HCl pH 7.4, 1% NP-40, 0.5% Na-deoxycholate, 0.1% SDS, 150mM NaCl, 2mM EDTA, 50Mm NaF) supplemented with protease inhibitor and phosphatase inhibitor cocktail (Gendeport, Barker, TX, USA).

5% non-fat milk in Tris-buffered saline pH 7.6 containing 0.1% Tween-20.

Overnight at 4°C.

1:3,000 dilution anti-mouse secondary antibody (Cell Signaling Technology, Cat# 7076) for 1 h at room temperature.

The antigen–antibody complexes were detected using West-Q Pico Dura ECL Solution (Gendeport, Barker, TX, USA).

Western-blotting to monitor TET2, p53, Atg16L1 and LC3 levels in p53-inducible HEK293T cells with or without depletion of the indicated autophagy-related genes. Please see Figure 4b in the article [1].