Western blot of the expressed missense TM mutants.

SDS-PAGE under nonreducing conditions and immunoblotting of cell lysates expressing each of the mutants or the wild-type TM were carried out. Immunodetection was performed with sheep polyclonal anti-TM antibody followed by HRP-conjugated antigoat antibody and ECL detection. The molecular weight markers were reduced and following electroblotting were detected separately with streptavidin-HRP conjugate.