Fwd: protein purification

Dr Engelbert Buxbaum via methods%40net.bio.net (by engelbert_buxbaum from hotmail.com)
Tue Aug 19 08:35:33 EST 2008


Am 11.08.2008, 23:50 Uhr, schrieb kamalaker nasani <agbiok4 from gmail.com>:

> I need to concentrate 66KDa size protein in my sample.(from plant coded  
> by
> transgene)
>
> Can any one give the suggestions regarding this
>
> What concentraion I have to go for Ammonium sulphate saturation and any
> other method is there to purify?

There is no good correlation between molecular mass and the AS  
concentration required for precipitation. You need to determine the  
maximal concentration at which your protein does not precipitate and the  
minimal conc for total precipitation in a series (say, every 10%). If you  
have an antibody against your protein, spot-blots of supernatants and  
precipitates are the way to go.

But for simple concentration (rather than purification) however I'd  
probably use ultrafiltration, not AS precipitation. Amicon pressure cells  
(say, with a 30 kDa membrane) do that neetly and are available for volumes  
 from a few ml to several l.


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