MBP versus GST...and the winner is...

bipin dalmia dalmiabk at phibred.com
Tue Apr 11 16:31:29 EST 1995


In article <Daniel.L.Burgess-0304951502120001 at 174.52.med.umich.edu> Dan
Burgess, Daniel.L.Burgess at um.cc.umich.edu writes:
>My questions are these:  Has anyone tried both?  Alternatively, if 
>you've had some experience with either, were you satisfied?  I'm not
>looking for the perfect system, but I would like my first attempt to
>have at least a 'snowballs chance in Michigan' of working.

it depends on what you'll be using the antibodies for. GSTs are
omni-present (like ubiquitin) but the antibodies to GST from one species
generally do not cross-react with GST from other species even though
there is high homology. i do not know about the presence in various
species for MBP. in either case, you can purify loads of 'native' GST or
MBP, immobilize them on a column (pierce sells kits) and subtract your
antibodies to get rid of the GST (or MBP) portion of the antibodies.

in my experience, GST has worked better than MBP, and thrombin cleavage
(i know you don't want to cleave it, but if you did) is much easier than
FXa cleavage.

bip



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