protein A tagged fusion protein - (Jun/07/2005 )
I have a fusion protein which is linked to protein by a TEV cleavage site. After digestion overnight with TEV protease, while it's obvious TEV did cleave my fusion protein, the problem is I cannot detect my protein on SDS gel (stained with Coomassie), but the protein A and TEV protease appear on my gel. Why is it that I cannot see the cleaved protein? I thought it may have been a problem with glycosylation but my protein is not very glycosylated. We all know that CHO's do not stain well. Another problem is that my cleaved protein runs at a MW very close to that of the TEV protease. I'm afraid TEV may be covering my protein, so I can never tell if I have the actual protein or not. Anyone have suggestions/solutions?
ur protein may be over-digested with TEV protease. use western blot to monitor the presence of ur cleaved protein