Combining gel filtration and electrophoresis
ben.long at yourfinger.anu.edu.au
Thu Feb 2 00:31:21 EST 2006
Daniel Käsmayr wrote:
>>> Is there a way to combine gel filtration with electrophoresis - i.e.
>>> separating proteins by size but inverse to that of a general SDS-PAGE?
>> 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.
Could you just run a normal gel and then turn it upside down? :-)
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