in- gel trypsin digestion - (Apr/20/2005 )
i tried to do an in-gel as well as in-solution digestion with trypsin but unfortunately i don´t see the desired peaks on the MALDI. i perform the digestion at 37 deg, overnight at an enzyme:substrate ratio of 1: 100. could anyone suggest where the problem can be. my protein is quite hydrophobic.
How do you stain your gel? is it by Sliver staining? if yes you need to check if the stain is mass spec comaptible, may be your protein is irreversibly cross linked to the gel and your not getting it in sloution after tryptic digestion? I hope these notes may help, good Luck!