FAQ: I see minimal cleavage of my glycoprotein after treatment with O-Glycoprotease. How can I optimize the reaction?

Tightly folded glycoproteins and/or glycoproteins with clustered O-glycosylated sites can be resistant to cleavage with proteases. In these cases, denaturing the substrate prior to digestion with O-Glycoprotease may be necessary for optimal digestion. In addition, longer incubation times (18 hours) and more enzyme may be necessary. However, please note that detergents and additional reagents must be removed from the reaction prior to the addition of O-Glycoprotease.