Hi, I'm having trouble in western blot for kinase (Ca2+/calmodulin-dependent protein kinase II ) and a phoshatase (calcineurin a) proteins. Those proteins don't appear in membrane. I already tested different protein concentration, extraction protocols (protease inhibitor cocktail SIGMA, direct lysis in laemmli buffer), different transfer conditions (overnight, 1h, 2h...). So, I would to know if exist something unique for detect that kind of proteins.
Thanks.