Hi, I have used tev protease for tag cleavage during dialysis. In my case always tev got precipitated not my protein. And cleavage was always complete. I havE checked on the sds - page as wel. On 2 May 2015 23:27, "Giulliana Rangel" <giulliana.ran...@gmail.com> wrote:
> Dear all, > > I'd like some help about my protein cause I've a lot of problems in > cleavage moment. In this step after aproximadately 30 minutes (37C) occur > precipitation almost 50% . > I tried control it: > - Protein diluition (no results) > - Cleavage 4C ( no cleavage) > -Modifying buffers: add 10% glycerol and 5% glucose (no crystallization) > - Add salt (1M - no results) > - Add serial tev (500ul in the first time and more 500ul in second time- > 37C) total precipitation > - Crystallization with 7 histag ( poor crystallization, no diffraction) > > Now I need to produce this protein with semet that became the protein more > hidrofobic, probably. > > So, If anyone could help me... > > Thanks in advance > > Giulliana Rangel >
