5'->3' InhibitACE

Andre Hamel hamel at cc.umanitoba.ca
Tue Oct 4 13:27:23 EST 1994


Has anyone compared effects of +/- DTT on activity of such RNase
inhibitors? ... literature supplied with InhibitACE from 5' -> 3' it is
MOST active in ABSENCE of DTT (they cite 10% drop in activity in presence
of 20 mM DTT). Inhibit-ACE is supplied in 150 mM NaCl, 2 mM KH2PO4, 8 mM
Na2HPO4, 3 mM KCl, pH 7.4 and 50% glycerol.

Their unit definition (1 U inhibits 90% of activity of 250 ng of RNase A) is
assayed in 30 uL reaction containing 40 ug yeast RNA, 250 ng RNase A, 25
mM Hepes-KOH, pH 7.6, 25 mM Tris-Cl, pH 7.5, 10 mM MgCl2, 50 mM KCl, 1 mM
spermidine, 1 mM EDTA, 5 mM DTT and Inhibit-ACE ... 37*C/2 hrs, analysed
by 1% agarose gel with EtBr staining.

all best,

                            ********************
Andre Hamel                                    email: hamel at cc.umanitoba.ca
Manitoba Government Veterinary Services          lab tel.: (204) 945-7630
545 University Crescent,                              FAX: (204) 945-8062 
Winnipeg, Manitoba, 
CANADA   R3T 5S6            ********************




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