Combining gel filtration and electrophoresis

Bean Long 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.
> 
> Dan
> 

Could you just run a normal gel and then turn it upside down? :-)


-- 
Bean

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