DTT "inhibitor" ?

BL benny at mrc-lmb.cam.ac.uk
Tue Dec 4 05:25:35 EST 2001

Is there any reagent that will specifically stop the -S-S- breaking (i.e.
reducing) ability of dithiothreitol, without affecting proteins otherwise?
I am trying to break the -S-S- bonds in some soluble proteins by 8M Urea
and 50mM DTT. However I need to get rid of the DTT completely before the
next step as a linker I use contains an -S-S- bond. The only way I can
think of is to use dialysis to get rid of the DTT. Is there a more
complete way?

On a related note, say, when I have dialysed the DTT against just 8M urea,
how quickly will the -S-S- bonds re-form, considering that it's in a
denaturing environment?

Thanks for any advice you can offer.

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