Ian McFarlane (imcfarlane at ninewells.dundee.ac.uk) wrote:
: Does anyone know of a quick and ideally easy way to purify sialoglycoproteins
: from serum. Is their an ion exchange method eg DEAE or a precipitation method,
: I seem to remember reading something about tungstante or vanadate or
: something. Basically what I am trying to achieve is removal of serum albumin
: friom the serum so I can look at the sialoglycoproteins by themselves.
: Thanks In Advance Ian Mc
You can remove serum albumin easily by using Affi-Gel Blue or CibaCron
Blue agarose. the blue dye is a ligand for albumin. Wash you column in 1M
Salt if you want and everything but the albumin will drop off (aalbumin
comes off in 0.5M KSCN). There is a Biochem J paper bu Gianazza and Arnaud
(Biochem J. 203 p637 (1982)) detailing the conditions required to
fractionate proteins. Alternativly use a combination of Jacalin agarose
and conA agarose. These should remove all biantennary N-linked
glycoproteins and mucin type glycoproteins from serum. You can thenm
re-pool these fractions and work o that. I would go witht he Cibacron
Blue agarose though.
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