avidin/streptavdin-biotin
Mr. M.J. Lush
mlush at hgmp.mrc.ac.uk
Mon Jul 22 07:17:01 EST 1996
In article <31EFC8A3.32A4 at uic.edu>, Keld Sorensen <KeldS at uic.edu> wrote:
>Streptavidin and avidin are both VERY hardy proteins.
>A little boiling won't hurt them and the biotin will stay.
>The avidin-biotin complex will withstand 3 M guanidine.
>Streptavidin will "live through" regular PCR protocols
>which usually are a minute at 94°C followed by lower
>but still high temps. A regular PCR protocol is up to
>40 cycles!!
>Hope this clarifies.
>Keld
This is true, but I understand that the boiling does sperate
the subunits and releases the biotin, but they will rapidly reassemble
and grab it back once normal temp is resumed. Boiling for 10 minutes
in 0.1% SDS will denature the protein and releace at least some of
the biotin. There are other ways of releasing biotin but they
are even more harsh!
--
Michael
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
NPC rights activist | Nameless Abominations are people too.
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