You mean the interested protein for the western blot? Is the protein membrane bound? Normal lysate protocols only work with secreted /soluble proteins. Membrane bound proteins usually requires something that destroys the bilayer in order to pull out the protein. I think RIPA is one of the most common?

If it's a surface protein... much easier to do flow.

Shen-An Hwang Thank you so much for your kind suggestion. My interest protein is a lipid raft component and should be bound to membrane to some extent. It's facing cytoplasmic not extracellular.

Dear I support the use of RIPA