processing of tissue for ELISA - (Oct/06/2008 )
I would like to develop an ELISA for an intracellular protein (in enterocytes).
Is there anyone who has a protocol for the processing of tissue for this purpose?
The protein is membrane associated.
I think immunocytochemistry may be more suitable.
Since the protein is intracellular (if it isn't a trans-membrane protein) I think ELISA might work better. You could probably sonicate the cells, centrifuge and pellet cell debris. Ultra-centrifuge the supernatant to pellet the protein fraction. Resuspend, dilute and run an ELISA.
Just a suggestion though. Might want a second opinion, and someone with a more detailed protocol.
I believe you are talking about developing an entire assay...making the abs to the specific protein and then using the ab to develop an elisa. Your question pertained to isolating the protein first...Do you already have antibodies/antisera or positive controls?
Can you elaborate?