Immunohistochemical analysis of mouse cochlear tissue labeling SLC26A5 with ab242128 at 1/50000 (0.011µg/ml) dilution.

The image is kindly provided by Dr. Zhiyong Liu, Institute of Neuroscience (ION), CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences (CAS), Shanghai, China.

Prestin, which is encoded by Slc26a5, is specifically expressed in inner ear cochlear outer hair cells (OHCs). Whole-mount prepared cochlear samples were blocked with solution containing 1% Triton X-100, and 5% BSA in 1x PBS for 1 h at room temperature, followed by primary antibody (1:50000) resolved in antibody solution containing 0.1% Triton X-100, and 5% BSA in 1x PBS at 4°C overnight. After 3 times 1x PBS wash, samples were stained with goat anti rabbit secondary antibody conjugated to Alex-Fluor^® fluorophores 488 at 1/500 dilution for 4 hrs at room temperature. After 3 times 1x PBS wash, samples were counter-stained with Hoechst 33342 to visualize nuclei. Samples were scanned with Nikon C2 confocal microscope. Counterstained with Hoechst.