Source for Factor Xa

Michael S Curtis (Michael_S_Curtis@ccmail.bms.com)
Mon, 24 Aug 1998 23:29:54 -0400

Can anyone recommend a source for Factor Xa? We are using it to cut
MBP fusion proteins. We have used material from NEB and Boehringer
Mannheim and have had moderate success cleaving the fusions. The
efficiency is poor and there is allot (>70%) of uncut fusion. We have
added additional enzyme, let it go longer (>24 Hours), tried
increasing temperature (RT, 30, and 37), added SDS (0.01, 0.05 and
0.1%); none of these gave improved cleavage. We are currently using
approximately 10ug of Factor Xa per 500ug fusion protein. We are going
through allot of Factor Xa and are interested in trying either a new
source (we need to buy some more enzyme) or a new protocol if anyone
has anything that isn't already available on the NEB web site.

Thanks for your help.

Mike


Michael Curtis
Bristol-Myers Squibb Company
and SUNY Health Science Center
Syracuse, NY