william.riordan at chirondiag.com
Mon Jul 20 12:22:22 EST 1998
Hi All -
I'm using a commercial prep (Sigma) of UDP-glucuronyltransferase
and , despite following some time-tested protocols - including one
provided by Sigma - I'm getting no activity. The company says no one
else has reported this problem.
Typical assay conditions are: Tris buffer (pH 8), with millimolar
quantities of MgCl2 and UDPGA, and milligram quantities of UDPGT. For
'acceptor' substrates, I've tried 4-nitrophenol and phenolphthalein,
also at millimolar concentrations. I've also tried various detergents,
as well as phospholipids, which are known to be important for the
activity of this enzyme. Still no luck.
If anyone has any suggestions/comments about the use of this
enyzme, I'd be very grateful. Experiences with Sigma's prep (the only
commercial source I'm aware of) would be especially welcome.
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