This is a knockout-validated antibody summary, based on the publication "β1-C121W Is Down But Not Out: Epilepsy-Associated Scn1b-C121W Results in a Deleterious Gain-of-Function", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: Na Channel β1 Subunit

Catalog number: 13950

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues surrounding Ala51 of human sodium channel β1 subunit protein. Reacts with human, mouse and rat. Suitable for western blot and immunoprecipitation.

Western blot

Mouse brain membrane extracts from WT and Scn1b−/− mice. Complete protease inhibitors (Roche Diagnostics) were added to all solutions at twice the recommended concentration to minimize protein degradation. Brains were dissected and homogenized in ice-cold Tris-EGTA (50 mm Tris, 10 mm EGTA, pH 8.0).

5% nonfat dry milk and 1% bovine serum albumin in TBST (0.1 m Tris-Cl, 0.5 m NaCl, pH 7.5, 0.1% Triton X-100).

1:3,000 dilution overnight at 4°C.

1:500 dilution HRP-conjugated goat anti-rabbit (Thermo Fisher Scientific) for 1 h at room temperature.

West Femto or West Dura reagent (Thermo Fisher Scientific, catalog #34095 or 34076) was used for detection. Immunoreactive signals were recorded using a Leica Odyssey Fc Imager.

Representative Western blots comparing anti-β1 immunoreactivity in Scn1b+/+, Scn1b-/- and Scn1bW/W mouse brain membrane preparations. α-tubulin was used as loading control. Please see Figure 2a in the article [1].

The article lists the correct clone number (but wrong catalog number) to indicate the antibody (from the author's feedback).