Enzyme Assay

Sabyasachi sabyab at yahoo.com
Sat Nov 4 12:27:03 EST 2000


Hi
I'm assaying NADPH-Cyt P450 Reductase & NADH-Cyt b5 reductase using
dichlorophenolindophenol as electron acceptor. I've found that most workers
have used Cyt c as electron acceptor. Am i doing anything wrong by using
DPIP? Also in a protocol for assaying quinone reductase or DT-Diaphorase I
see that the the protocol used is essentially similar to the method i'm
using. All protocols use an artificial electron acceptor, similar buffers at
similar molarity & pH, & either BSA or tween. Except that the latter to
enzyme assays use dicumarol as an inhibitor, it is supposed to be specific
for DT-Diaphorase &/or quinone reductase. What I want to know is are all
these enzymes the same?
I'll be grateful for any information.
Thanks
Sabya







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