Chemicon protein concentrate kit - what's in it?

Frank O. Fackelmayer Frank.Fackelmayer at uni-konstanz.de
Wed Aug 25 02:11:42 EST 1999



Peter wrote:

> In article <RNA.world-2408991650140001 at expmac19.uni-muenster.de>,
> RNA.world at uni-muenster.de (Juergen Brosius) wrote:
>
> > Chemicon offers a protein-concentrate kit using a "Universal Protein
> > Precipitating Agent (UPPA)" and suspension buffer for precip. of proteins
> > out of solutions as low as 1 ug/ml.  Any idea what might be in it?
>
> I could do even better.  I have sucessfully gotten 4 ng/ml out of solution
> and all my reagents can be gotten from our general supplies(Phenol and
> Ether)
>
> regards,
> Peter

Hi Juergen, Hi Peter,
I don´t have an answer to the original question, but I´d like to add a thought
from the protein side. Of course you can quantitatively recover proteins from
dilute solutions with quite a number of methods (Phenol/Ether,
Methanol/chloroform (that´s the one I prefer), aceton....), so that´s not the
problem really. The problem from the protein side is that these methods
usually work by denaturing the proteins and forcing them to aggregate. That is
perfectly ok when you want to concentrate the sample for SDS PAGE, for
example, but is not at all useful if you want to do something functional with
the protein. So the problem is: How can I recover protein from dilute solution
but retain activity? The answer to that question is far more difficult to find
and also variable with the protein of interest. The classical (and usually
good) method is precipitation with ammonium sulfate, but other substances have
also been used successfully (PEG, Polymin...).

Frank




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