2 replies to this topic

[molecular_medicine]

Hi all,

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!!!

[Rsm]

Hi there,

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...

Best,

Minna

[bob1]

Antibodies are also relatively large, getting them into the cell is likely to be difficult.