westernblot problem! - membrane protein! (Feb/12/2006 )

Hi,
I have a new antibody,and should find the molecular weight of antigen,I tried westernblot several times with SDS,Native gel but I didn't have any bands,I think ,it's a membrane protein ,I used some membrane protein extraction kit(PIERCE)but nothing!!
this membrane protein can not run in SDS and Native gel !!what's your idea ??what can I do?do you have a protocol for lysis of membrane protein ??is there any special gel for runing membrane protein?please help me??

-soha-

Did you boil your protein prior to loading it on the gel?

Membrane proteins in my experience do not cross react well with the antibody if you have boiled them.

-pBluescript-

QUOTE (pBluescript @ Feb 14 2006, 02:21 AM)

Did you boil your protein prior to loading it on the gel?

Membrane proteins in my experience do not cross react well with the antibody if you have boiled them.

For native gel I didn't boil but for SDS gel I did,I think this is a cell membrane protein!

-soha-