reduction using DTT
ballal_sv at hotmail.com
Tue Jul 29 10:40:53 EST 2003
As per our experience 10 mM DTT should be enough. Incubation for 30
min. at RT seems to do the job, though I am not sure if the no. of
di-sulphide bonds in a protein should effect, but I guess not.
Please ensure that the DTT is freshly prepared or the stock is frozen
all the time or else DTT being volatile will vanish very soon.
Unfortunately DTT absorbs well at 280 (and may be at 260 too) so it
will interfere with standard UV spectroscopy.
Best of luck
biplabbose at hotmail.com wrote in message news:<20030727112736.4770.qmail at ww02.hostica.com>...
> I want to performe stability study of a protein using spectroscopic
methods like CD, fluorescence. As the protein of interest have
disulfide bond, I want to reduce it using DTT, so that we can
understand the importance of the disuphide bond in stability of the
protein. What is the usual protocol for reduction of protein for such
studies? Will 10mM-20mM DTT and incubation at room tem for 2 hr / or
overnight incubation at 4 degree, do the job? Does DTT interfare in
> Thanking you
> Biplab Bose
> New Delhi
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