C-terminal Analysis Protocol

ez027854 at othello.ucdavis.edu ez027854 at othello.ucdavis.edu
Thu May 20 14:17:31 EST 1993


To all those folks who replied with helpful suggestion, Thankyou. :)

After examining the suggestions received and the literature closely,
I decided that the standard carboxy peptidase y approach would yield the
best initial results.  I came across a few chemical approaches that were
recently published utilizing thiohydantoin derivatives but found (through
discussion with those more familiar with thiohydantoin chemistry) that
this method is still fairly "dirty".

This leads me to my next question, which is...  Now that I have decided to
use carboxy peptidase y which company is the best provider of this enzyme?
I have seen it from both Sigma and Boeringer Mannheim, although BM is the
only producer I have encountered so far that makes a sequencing grade enzyme.
Is it necessary to purchase the sequencing grade as opposed to the analytical
grade?  Is contaminating proteolytic activity a significant problem in
analytical grade preparation of this enzyme?

And... I am considering doing the amino acid analysis myself on a C18 reverse
phase column.  Does anybody out there have a protocol or reference that I
may turn to for more precise guidance in this area?

					Thankyou
					Mike Oda
					ez027854 at hamlet.ucdavis.edu



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