Hi Folks,
We are trying to immpt. a yeast mitochondrial membrane protein which has
been tagged with the HA epitope. In our hands, the only way we can access
the epitope is to spheroplast, solubilize in 1% SDS and dilute the
extracts to 0.1% SDS before adding antibody. As far as antibodies, we
seem to get better recovery if we use the HA.11 monoclonal from BabCo
rather than the 12CA5 monoclonal. The trouble with this is that we are
getting background that we are having trouble eliminating. Has anyone
tried the BabCo polyclonal antibody to the HA epitope for immpt. and is
the background better? Any suggestions or experience here will be
appreciated.
Mike Forte
--
Mike Forte
forte at ohsu.edu or
forte at mtwo.com
