VOLATILE BUFFERS FOR COLUMN CHROMATOGRAPHY

[NICHOLAS THEODORAKIS]

In article <092ellbj.5 at chiron.wits.ac.za>,
Bruce James Ella <092ellbj at chiron.wits.ac.za> wrote:
>I am fractionating the low molecular weight proteins in serum on a G50 
>Sephadex column. After which I need to freeze dry the 6 fractions but remove 
>the salt concentration such that the sample can be added to cell medium. 
>I figured that elution of the samples from the column with a volatile 
>buffer, eg ammonium formate or carbonate, would be the answer. Can any one 
>help?

It sounds like you already know the answer.  This should work, provided 
your protein of interest is stable in your buffer and to lyophilization.  
Many years ago, when I was a tech., the lab I worked in did the final 
step of PDGF purification by chromatography over BioGel P-100 in 1 M 
acetic acid; we then pooled and lyophilized the active fraction as you 
describe.

					Nick

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			Nick Theodorakis
			ntheo at welchlink.welch.jhu.edu
			Johns Hopkins Medical School, Baltimore, MD