How to dispose beta-Mercaptoethanol out of solutions

[Nick Theodorakis]

Sebastian Gruenberg wrote:
> Hi members,
>
> I have an urgent question for the forum.
>
> I'm doing KMnO4-footprinting experiments and unfortunately, BME
> disturbes the KMnO4 reaction (it is also used to quench the reaction
> later). Since there is BME in the buffers of the proteins used in my
> reactions, I have to get rid of almost all the BME in the protein
> solutions.
>
> Does anybody now how to do this without damaging the sensitive
> proteins?


Iodoacetic acid or (better yet) iodoacetamide will alkylate free
sulfydryls. Add a slight molar excess (e.g., 2-fold) over free
sulfhydryls and incubate in the dark for about 30 min. (Assuming this
will not affect the permanganate, which I don't know). Of course, if
your protein has free solvent-accessible -SH than they will be
alkylated as well.

Nick

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Nick Theodorakis
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