Paper bridge method for 2D electrophoresis

Elias Klein e0klei01 at athena.Louisville.edu
Tue Oct 9 14:20:46 EST 2001


I read the paper you referred to in your note.  These authors used a
narrow pI range IEF strip (4.5 to 7.5) and avoided the albumin
interference by running it off into the buffer.  How did you reduce the
albumin concentrations:
    For the volumes you are going to use I would suggest using a
centricone ultrafilter.  You can spin the sample and remove water and
microsolutes through the membrane. Amicon/Millipore as well as other
companies (Rainer,  Pierce) make such devices with different mol. wt
cut-offs and retention volumes.  Good luck.  E.K.

Anne-Françoise Marchand wrote:

> Hello,
> I'm performing an electrophoresis 2D on plasma with the paper bridge
> method.
> My human plasma protein concentration, after extraction of albumin, is
> about 10ug/ul. We can load a maximum of 500ul on a paper bridge of 2.5
> cm in lengt.  So, we can load 200ul of plasma in 250 ul sample
> buffer.  That means 2 mg of proteins.  Or, in an article of
> Sabounchi-Schutt (Electrophoresis 2000), I red that we can load up
> to7.5mg of protein.  Would you please explain how concentrate the
> sample. I tried 1 hour in a speed vak but it doesn't work, the gel was
> bad.
> Thank you for your help.
> Anne-Françoise Marchand
> anne-francoise.marchand at umh.ac.be
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