Proteins were electrophoretically separated on 7.5 or 10% SDS-PAGE (Bio-Rad, Hercules, CA) using Tris-glycine sodium dodecyl sulfate (SDS) buffer (Bio-Rad) at a constant voltage of 180 V. The electrophoresis buffer for the native gels did
not contain SDS. Protein bands were visualized by staining with Coomassie brilliant blue R-250. For Western blot analysis, protein was transferred onto nitrocellulose membranes (Bio-Rad) in 10 mM CAPS, 0.07% SDS buffer at 30 V overnight.
The TRPM8 protein was detected with anti-Myc-IgG antibodies.