serum protein delipidation question

Brendan P Hughes Brendan at bphughes.demon.co.uk
Wed Mar 15 15:35:34 EST 1995


In article <3k506i$n1n at dove.nist.gov> dmb618 at enh.nist.gov "D.M. Bunk" writes:

> Does anyone have any "tried-and-true" methods (or 
> references to methods) for defatting serum proteins?
> 
> I've been trying to isolate albumin from serum on a 
> microscale using antibody affinity chromatography.  
> While I have been able to extract the albumin, the 
> albumin retains its complement of bound small 
> lipophilic molecules (which interfere with the 
> quantitation I ultimately want to do).
> 
> I have tried to defat the extracted albumin using the 
> standard free fatty acid extraction solvent of 
> chloroform/MeOH (2:1).  The problem here is that the 
> MeOH precipitates the protein which settles on the 
> chloroform-water interface, making it difficult to 
> recover.
> 
> Does anyone have any suggestions?
> 
> -David
> 
You might want to try Lipidex.  This is a strong fatty acid binder and was used
to extract fatty acids from Fatty acid binding proteins to allow enzymatic
peptide mapping.  I can't remember the exact paper but if you do a search for
J.I Gordon in the literature with key words fatty acid binding, you should 
find a J Biol Chem paper describing the procedure.  Published between 1986-1988.
Sorry to be so vague, but I've been out of the area for a long time.

-- 
Brendan P Hughes



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