TCA precipitation of proteins

Mark Bowen mb at laplace.csb.yale.edu
Mon Jul 17 17:00:08 EST 2000


If you are using affinity chromatography why can't you bind your protein
to a small amount of affinity matrix and step elute it into a very small
volume.  I work with membrane proteins and keep the His tag on for
concentration and detergent exchange.

Concentration does not work very well for detergent containing samples.
Detergent micelles are often <90kDa and will not pass through membranes
(dialysis or concentrator).  Monomers will pass through but micelles
take a long time to dissociate and pass though as monomers.  You will
end up increasing your detergent concentration along with your protein
in most concentrating schemes.

--
Mark Bowen







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