This is a knockout-validated antibody summary, based on the publication "One-step generation of triple knockout CHO cell lines using CRISPR/Cas9 and fluorescent enrichment", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Rabbit polyclonal IgG

Company: Sigma

Antibody: Bak

Catalog number: B5897

Summary: Rabbit polyclonal IgG against a synthetic peptide corresponding to the N-terminal of human Bak (amino acids 23-38 with C-terminally added lysine). Recognizes human Bak by immunoblotting and immunohistochemistry.

Western blot

Wild type, BAK/BAX double knockout cell line and FUT8/BAK/BAX triple knockout CHO‐S cell lines. Lysis buffer (10 mM Tris‐HCl, pH 7.4, 100 mM NaCl, 2.5 mM MgCl2, 0.5% Triton X‐100, supplemented with 1x Roche protease inhibitor cocktail).

5% dry skimmed milk diluted in 1x PBS and 0.1% Tween20 for 1h.

1:2,000 dilutions overnight.

1:5,000 dilution HRP‐conjugated donkey anti‐rabbit IgG secondary antibody for 50 min.

Amersham ECL detection system (GE Healthcare Life Sciences, Buckinghamshire, UK).

Please see Figure 2b in the article [1].

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.