This is a knockout-validated antibody summary, based on the publication "ABHD5 interacts with BECN1 to regulate autophagy and tumorigenesis of colon cancer independent of PNPLA2", as cited below [1], and "Western diet induces severe nonalcoholic steatohepatitis, ductular reaction, and hepatic fibrosis in liver CGI-58 knockout mice" for western blot knockout validation (figure 4b) [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Mouse monoclonal IgG2a Kappa

Company: Abnova

Antibody: ABHD5

Catalog number: H00051099-M01

Summary: Mouse monoclonal IgG2a Kappa antibody raised against full length recombinant ABHD5. Reacts with human and rat. Suitable for western blot, immunohistochemistry (paraffin), and ELISA.

Western blot

ABHD5+/+ (WT) and ABHD5-/- (KO) CCD841CON cells. Cells were lysed on ice for 15 min in RIPA buffer supplemented with a protease and phosphatase inhibitor cocktail (Sigma, P8340 and P2850), followed by brief sonication.

5% nonfat milk dissolved in PBS-Tween 20 (0.1%; Beyotime Biotechnology, ST825) solution for 1 h.

In PBS-Tween 20 containing 1% BSA (Biosharp, BS043E) overnight at 4°C.

HRP-conjugated secondary antibody.

Enhanced Chemiluminescence (ECL) detection reagents (PerkinElmer, NEL105001EA). The signals were captured using a Bio-Rad ChemiDoc MP System (170-8280).

Western blots of autophagy-related proteins (LC3-I, LC3-II, SQSTM1) in ABHD5+/+ (WT) and ABHD5-/- (KO) CCD841CON cells at 24 h following exposure to PBS, EBSS or EBSS+rapamycin (100 nM). Please see Figure 1k in the article [1].

The same clone (1F3) is sold as OriGene AM31371PU-N; MilliporeSigma WH0051099M1; LifeSpan Biosciences LS-B5751; Abnova H00051099-M01.