Deglycosylation kinetics
PBCV (5 microliter aliquots, dried overnight) was deglycosylated with 5 microliters metacresol and 20 microliters of trifluoromethanesulfonic acid at either 0C or RT. Samples were neutralized with 3 volumes of N-ethylmorpholine and precipitated by adding 5 volumes of ethanol, 2 microliters of 1% Ficoll and incubation at -20C for 30 min.
Sample on the far right was deglycosylated 2hrs at RT, but w/o cresol (scavanger). This samples was much darker than the others prior to acetone precipitation. Obviously the scavanger is serving a useful function.
Half an hour at 0C appears sufficient for PBCV deglycosylation.
