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


Dear netters,
    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 
help.  

Sincerely,
John Marcus



John Marcus
Cooperative Research Centre for Tropical Plant Pathology
5th Level John Hines Building
University of Queensland
Brisbane, QLD 4072
Australia
Marcus at tpp.uq.oz.au



More information about the Proteins mailing list