We were hoping to use the b-galactosidase moiety of our fusion protein
as an epitope tag. We purchased monoclonal antibody against b-gal from
Promega, but found on Westerns about 6 extra bands even with native
b-galactosidase. The Promega tech says that the protein is heat labile,
and is being degraded in the loading buffer during heating prior to loading.
Apparently they have only characterized the Ab with purified protein, and
had nothing useful to suggest about how to solve the problem in cell
extracts (we are using E. coli).
Has anyone else tried a similar approach, and have any useful
experience to share. Do we give up?
Patricia L. Foster
Boston University School of Medicine
Boston, MA USA
pfoster at bu.edu