This is a knockout-validated antibody summary, based on the publication "The adherens junctions control susceptibility to Staphylococcus aureus α-toxin", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Rabbit polyclonal

Company: Sigma

Antibody: Afadin

Catalog number: A0349

Summary: Rabbit polyclonal against a synthetic peptide sequence corresponding to amino acid residues 1814-1829 of rat afadin. Reacts with human, canine, mouse, and rat. Suitable for western blot, immunohistochemistry (frozen sections), immunofluorescence and microarray.

Western blot

WT and Afadin-knockout HAP1 cells. Cell pellets were lysed with hot Laemmli SDS sample buffer containing 5% (vol/vol) β-mercaptoethanol and boiled for 10 min.

PBS buffer containing 5% (wt/vol) nonfat milk and 0.01% Tween-20 for 1 h at room temperature.

1:2,000 dilution in blocking buffer overnight at 4 °C.

1:5,000 dilution HRP-conjugated anti-rabbit secondary antibodies (GeneTex) in blocking buffer for 1 h at room temperature.

West Pico and Extended Duration chemiluminescence peroxide solutions (Thermo) and film exposure.

Please see Figure S1 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.