This is a knockout-validated antibody summary, based on the publication "Deafness induced by Connexin 26 (GJB2) deficiency is not determined by endocochlear potential (EP) reduction but is associated with cochlear developmental disorders", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Mouse monoclonal IgG2a, kappa

Company: Invitrogen

Antibody: Connexin 26

Catalog number: 33-5800

Summary: Mouse monoclonal IgG2a, kappa against a 13 amino acid synthetic peptide derived from the C-terminus of the mouse Connexin 26. Reacts with mouse and rat, and has been published to react also with human, chicken, and guinea pig. Suitable for western blot, immunohistochemistry and ELISA.

Immunohistochemistry

WT and Cx26-cKO mouse cochlear sections. Cryostat cross-sections were fixed with

4% paraformaldehyde in 0.1 M PBS (pH 7.4) for 30 min.

10% goat serum and 1% BSA in the PBS (blocking solution) containing 0.1% Triton X-100 for 30 min at room temperature.

1: 400 dilution in blocking solution at 4°C overnight.

1:600 dilution of secondary Alexa Fluor 488 conjugated antibodies (Invitrogen, CA) in blocking solution at room temperature for 1 hr.

Fluorescence microscope (Nikon T2000) or confocal microscope (Leica TCS SP2).

Immunofluorescent staining for Cx26 (green) and Cx30 (red) in the cochlea for injection of 4-hydroxytamoxifen (4-HTMA) at different postnatal ages to delete Cx26. No intense Cx26 labeling in the cochlea is visible in Cx26 conditional KO mice. Mice were P30–45 old. Scale bar = 50 μm. Please see Figure 3 in the article [1].