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Dear Michal, the glycosylation in pichia is notoriously heterogeneous. As a result, endo H will stop cleaving at different branch-lengths and you end up with a mixture of truncated glycans. You could use a glycosylation inhibitor such as tunicamycin. Also, a modified strain of pichia was reported to have homogeneous glycosylation: Production of Complex Human Glycoproteins in Yeast (2003) Science 301, pp 1244-1246 cheers, Rob Meijers Dana Farber Cancer Institute --- Michal Harel <[EMAIL PROTECTED]> wrote: > *** For details on how to be removed from this list > visit the *** > *** CCP4 home page http://www.ccp4.ac.uk > *** > > > > > > > > We are expressing a cell adhesion protein which is > very > > heterogeneous in it's pI in Pichia. After > Deglycosylation by Endo H or > > by Endo F1 we > > can see 3 bands. Can anyone suggest what is the > reason for this > > heterogeneity? > > > > > >> > ------------------------------------------------------------------------ > Dr. Michal Harel > Dept. of Structural Biology > [EMAIL PROTECTED] > Weizmann Inst. of Science tel: > 972-8-9342647 > Rehovot 76100, ISRAEL fax: > 972-8-9344159 > ------------------------------------------------------------------------ > > __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
