I wish to a stain a HA-Tag protein by IHC from cell-lines implanted subcutaneously in mice. The protein is very unstable by nature and expressed at low levels. Is there any good well-tested anti-HA tag antibody for IHC of such proteins?
Any recommendation and direction to specific protocol is highly appreciated. Thanks.
Since you will be chemically fixing the sample before bindng, your unstable protein should not get degraded. Any commerical HA antibody that can immunopreciptate your protein should work well for IHC. As for the protocol, I guess the bigger problem will be figuring out the dilution to stain your sparsely expressed protein without significant background. Try adding 1% BSA or your preferred blocker in your binding buffer with the antibody. And a non immune IgG control should help to identify any background non specific binding in your samples. Hope this helps.
