Combining gel filtration and electrophoresis
daniel.kaesmayr at ch.tum.de
Wed Feb 1 12:30:46 EST 2006
>> Is there a way to combine gel filtration with electrophoresis - i.e.
>> separating proteins by size but inverse to that of a general SDS-
> Also not sure I understand the question but if it helps there are
> electrophoresis systems which enable elution of proteins for
> purification etc. BioRad sell a system with a cylinder of gel
> which is
> connected to a fraction collector.
What I would like to have is a system that uses electric charge to
drive particles, but also a matrix that retards smaller particles
much more than larger ones; resulting in something like an inverse-
PAGE, where larger particles migrate faster than smaller ones.
The substances that need to be characterized are star polymers /
brush polymers that have multiple functional groups (amine or
carboxylic acids) which are used to create peptide bonds to make
conjugates with DNA and/or Proteins.
What would also help is some sort of modification I could do to PAGE
that would help me get my star-shaped polymers (and their conjugates
with proteins or DNA) into the gel. Right now it seems like I either
do not get them to penetrate the gel matrix (I see only some residues
on the gel pocket's outside and just a few mm inside the gel - these
are 3% gels already and ver un-handlable) or they are not stainable
with a silver stain
From my syntheses I know the polymers can be analyzed by GPC/SEC -
but I don't have enough material for this anymore - DNA is very
expensive if you are talking about mg amounts.
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