Removal of TCEP

Wolfgang Schechinger via methods%40net.bio.net (by glybirne from gmail.com)
Mon Feb 20 03:26:29 EST 2012


Dear experts,

I would like to use TCEP (Tris-(2-carboxyethyl)-phosphine-
hydrochloride, https://en.wikipedia.org/wiki/TCEP) for reducing
disulfide bonds of a protein I want to couple to a maleimide
containing spacer. The advantage of TCEP is that it doesn't react with
maleimides, in contrast to BME or DTT, so the usual protocols do not
require/provide for quenching of excess TCEP.

In the next step, I would like to couple this protein to another
protein where leftovers of TCEP will harm the disulfide bonds. Is
there any way to quench excess TCEP? Passing through a column of
immobilized eg BSA comes into my mind. Any better ideas?

Thanks!

Wo





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