This is a knockout-validated antibody summary, based on publications "Pharmacological modulators of autophagy activate a parallel noncanonical pathway driving unconventional LC3 lipidation" (see below) [1], "The WD40 domain of ATG16L1 is required for its non‐canonical role in lipidation of LC3 at single membranes" (Western blot, see Figure 2b, 2c, 2d in the article) [2], "p53-dependent autophagic degradation of TET2 modulates cancer therapeutic resistance" (Western blot, see figure 4b in the article) [3], and "SPATA33 is an autophagy mediator for cargo selectivity in germline mitophagy" for western blot knockout validation (figure 4b) [4]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Representative western blots of ATG16L1 and GAPDH (loading control) in wild-type and ATG16L1−/− MCF10A GFP-LC3 cells treated as indicated. From [1].

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: Atg16L1

Catalog number: 8089

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues near the amino terminus of human Atg16L1 protein. Reacts with human, mouse, and rat. It is predicted to react based on 100% sequence homology with monkey. Suitable for western blot, immunoprecipitation and immunofluorescence/immunocytochemistry.

Western blot

Wild-type and ATG16L1−/− MCF10A cells. Cells were scraped into ice-cold RIPA buffer (150 mM NaCl, 50 mM Tris-HCl, pH 7.4, 1 mM EDTA, 1% Triton X-100 (Sigma, T8787), 0.1% SDS (Sigma, L3771), 0.5% sodium deoxycholate (Sigma, D6750) and lysed on ice for 10 min.

5% BSA in TBS-T for 1 h at room temperature.

1:1,000 dilution in blocking buffer overnight at 4°C.

Horseradish peroxidase-conjugated secondary antibody (Cell Signaling Technology, 7074S).

Enhanced chemiluminescence (GE Healthcare Life Sciences, RPN2209).