Western Blot Antibodies for inhibiting protein activity?? - (Mar/14/2010 )
I was wondering if we can use the antibodies that are used in western blotting to block the protein in cell culture and study its effects?
I want to inhibit a protein of interest, so i was thinking to use antibodies used in western blots to block it and then do a proliferation assay (or other assays) on it.
Please tell me if this is possible.. Many Thanks!!!
Most antibodies do not block protein function. They bind to other than the active parts of the protein. I know, however, that there's an antagonistic c-Kit antibody (Ack2), that blocks c-Kit signaling in vivo and in vitro (Disclaimer: our lab gets royalties for Ack2).
I'd recommend to check the literature carefully if there's any blocking antibody described for your protein. If it's a ligand/receptor pathway, you could also consider creating an Fc chimeric protein. But I'm not an expert for that...
Antibodies are also relatively large, getting them into the cell is likely to be difficult.