Immunodetection techniques are based on ... it may be a good idea to run a western blot first to make sure the antibody detects the correct protein and is not creating off-target signals.

For western-blot analysis, proteins from cultured RPE were electrophoresed on 12.5% SDS-polyacrylamide gel and then electrotransferred to a nitrocellulose membrane. We carried out immunodetection ...