Antibody Purification

Andrei Popov andrei.popov at SPAM.bbsrc.ac.uk
Wed Aug 27 09:51:46 EST 1997


Chris Weir wrote:
> 
> Hi all,
>   We have an IgM antibody which is difficult to purify from ascites!
> The problem is that it falls out of solution very easily. Have tried
> peg purification which usually is an effective method (for IgM's) but
> the final pellet wouldn't go back into solution ( in carbonate
> buffer). So I had to use a tris buffer to get it into solution.
> I then passed it through a column (equilibrated with Carb-buffer) but
> lost a lot of antibody in this step! and then conjugated what
> antibody was in solution. The trouble is that this conjugate is
> now precipitating out! Any suggestions
> 
> Chris Weir
> Macquarie University
> Sydney Australia











Salt is the answer (NaCl:-)

Carbonate buffer for ABs is an anachronism from the
Stone Age of immunochemistry:-)



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