Desalting small basic proteins on Sephadex G-10
MARCUS, DR J.
J.Marcus at botany.uq.oz.au
Tue Mar 21 01:09:52 EST 1995
I am using a Sephadex G-10 Column to desalt small basic
proteins. I would like to avoid tedious steps since I have
many fractions to desalt. Ideally, we would like to use a
volatile buffer system so that we are able to concentrate
our samples after desalting. I posted a message a few
months back and received many suggestions. Thank you all
for your help!
The most common suggestion was to use a 1% acetic acid
solution. This can easily be dried down afterwards to get
rid of most of the acetic acid.
Another suggestion was to use reversed phase to
desalt. The protein will elute in acetonitrile/TFA which
are both volatile.
It seems by using multiple drying/resuspension cycles
we are able to get rid of enough of the TFA for it not to
be a problem in our bioassays. The same would apply to
acetic acid. The problem is that all these drying down
cycles are TEDIOUS.
Has anyone ever tried ethanolamine as a volatile buffer
to use in conjuction with deslating on Sephadex G-10? Would it be a
problem for the vaccuum pumps? Thanks in advance for any
Cooperative Research Centre for Tropical Plant Pathology
5th Level John Hines Building
University of Queensland
Brisbane, QLD 4072
Marcus at tpp.uq.oz.au
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