protein isolation - (Feb/21/2002 )
how to isolate membrane protein from mammalian cells( t cells), and how to perform westren blotting to show this is expected protein. please any body suggest me the details protocol regarding this.
Thanks in advance
as first, you have to disrupt cells by sonication in an appropriate lysis buffer and second to separate plasma membrane on a sucrose gradient by ultra-centrifugation.
After this step, proteins are separated on PAGE for western blotting.
I send you by e-mail a PDF paper which describe this procedure for our protein of interest.