Luciferase Problems?

Clemens Suter-Crazzolara un691cs at genius.embnet.dkfz-heidelberg.de
Wed Nov 2 03:49:03 EST 1994


> 
> Hi, I was wondering if you could elaborate on this reply.  I have
> Hi, I was wondering if you could elaborate on this reply, I have never heard of this problem, ie: not being able to quantify it.
> Thanks,
> Sherri
> > > Hello netters, 
> > do only thing that comes to my mind is that luciferase is almost impossible
> > to quantify, even if you have a luminometer. Sorry, no exp. with beta-
> > gal
> > 
> > clemens, heidelberg
 
 unlike other enzymes, luciferase stays bound to D-luciferin (substrate).
 Thus, the amount of active luciferase is rapdily depleted from the 
 reaction mixture. To reduce this binding, I beleive acetyl-coA is added
 to the reaction; this allows to some extend an uncoupling of enzyme and
 substrate/product. 
 Generally, with luciferase, you get a peak flash in the first 0.1 secs
 or so, then the signal goes down rapidly. This is, as you may understamd,
 not ideal if you want to measure enzyme activity in the linear range...

 clemens, heidelberg




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