This is a knockout-validated antibody summary, based on the publications "Loss of PTEN Facilitates Rosiglitazone-Mediated Enhancement of Platinum(IV) Complex LA-12-Induced Apoptosis in Colon Cancer Cells" [1] and "BAX and BAK1 are dispensable for ABT-737-induced dissociation of the BCL2-BECN1 complex and autophagy" [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

[enlarge]

Figure 1. Western blotting of Bax protein in HCT116 wt and Bax/- cells. β-actin served as loading control. From [1].

Rabbit polyclonal

Company: Cell Signaling Technology

Antibody: Bax

Catalog number: 2772

Summary: Rabbit polyclonal produced by immunizing animals with a synthetic peptide corresponding to the amino-terminal residues of human Bax. Recommended for the detection of human, mouse, rat, and monkey by western blotting and immunoprecipitation.

Western blot

Human colon adenocarcinoma cell lines HCT116 wt (p53+/+, Bax+/-, Chk2+/+, PTEN+/+) and Bax-/-. Cells were lysed in 1% SDS lysis buffer containing protease inhibitor cocktail (#P2714, Sigma–Aldrich) or Protease Inhibitor Cocktail Set I (#5391313, Calbiochem, Merck Millipore, Bedford, MA, USA), for phosphoprotein detection NaVO4 and NaF were added to the lysis solution.

5% non-fat milk or BSA for 1 h at RT.

1:1,000 dilution in 5% non-fat milk at 4°C overnight.

anti-rabbit IgG antibody (1:3,000, #NA934) (Amersham Biosciences, Piscataway, NJ, USA) for 1 h at RT.

Immobilon Western Chemiluminescent HRP Substrate (#WBKLS0500, Merck Millipore, Bedford, MA, USA).

Western blot

Lysates of wild-type and BAX knockout HCT116 cells.

Lysates of wild-type and BAX knockout mouse embryonic fibroblasts.

[enlarge]

Figure 2. From Figure 1 in [2]. Western blot of lysates of wild-type and BAX knockout cells probed with anti-BAX antibody. GAPDH serves as a loading control.

TBS containing 5% milk powder and 0.05% Tween 20.

Anti-BAX antibody diluted 1:1000.

HRP-conjugated secondary antibody (Southern Biotech).

SuperSignal West Pico chemiluminescent reagent (ThermoFisher Scientific).

If the antibody described in this summary is a polyclonal antibody, since polyclonal antibodies are of limited quantity, please inquire the supplier whether any current polyclonal antibody with the same catalog number is exactly the same as the one described in this summary. Sometimes, different bleeds or different animals are used, usually with a different lot number. In such cases, the result in this summary may not apply to the new antibody with the same catalog number.