This is a knockout-validated antibody summary, based on publications "mTORC1 inhibition in cancer cells protects from glutaminolysis-mediated apoptosis during nutrient limitation", as discussed below [1], "XIAP facilitates breast and colon carcinoma growth via promotion of p62 depletion through ubiquitination-dependent proteasomal degradation" [2] (western blot, see figure 4b in the article), "Autophagy deficiency promotes triple-negative breast cancer resistance to T cell-mediated cytotoxicity by blocking tenascin-C degradation" for western blot knockout validation (figure 3b) [3]. This antibody was also validated for Western blot in the article "Knockout of autophagy gene, ATG5 in mice vaginal cells abrogates cytokine response and pathogen clearance during vaginal infection of Candida albicans" (see figure 1) [4], "TRIM14 Promotes Noncanonical NF-κB Activation by Modulating p100/p52 Stability via Selective Autophagy" for western blot knockout validation (figure 3a) [5], "Metformin activates chaperone-mediated autophagy and improves disease pathologies in an Alzheimer disease mouse model" for western blot knockout validation (figure s2i) [6], "Group III phospholipase A2 downregulation attenuated survival and metastasis in ovarian cancer and promotes chemo-sensitization" for western blot knockout validation (figure 6h) [7], and "Autophagy core protein ATG5 is required for elongating spermatid development, sperm individualization and normal fertility in male mice" for western blot knockout validation (figure 2d) [8]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Western blot analysis of ATG5 in WT and ATG5−/− MEFs. From [1].

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: Atg5

Catalog number: 12994

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues surrounding Leu265 of human Atg5 protein. Reacts with human, mouse, and rat. It is predicted to react based on 100% sequence homology with monkey, Xenopus, dog, and pig. Suitable for western blot and immunoprecipitation.

Western blot

WT and ATG5−/− MEFs. Cells were lysed with RIPA buffer containing a cocktail of protease inhibitor (P8340 Sigma), inhibitors of phosphatases (P0044 and P5726 Sigma) and PMSF 1 mM.

1:1,000 dilution.

1:1,000 dilution anti-rabbit (Cell Signaling Technology, #7074).

Membranes were imaged using the ChemiDoc MP imager (Bio-Rad).