Hi,
Thanks to everyone for the suggestions I've gotten. To answer some
questions, the protein I'm expressing is 6x-His tagged at the N-terminus,
and the Ab I'm using is a polyclonal against the protein rather than the
tag itself. Silver stained gels shows many more bands than my Westerns do.
Purification of the protein on two different resins, one Ni-NTA and the
other Talon, produced similar multi-banded results. It is possible that
the protein is expressed in a cell-cycle dependent fashion so it may be
degraded as the cells pass through certain stages of the cycle. If this is
the case, I suppose that using any combination of protease inhibitors will
be futile. Any thoughts?
Once again, thanks for the advice,
Victor
