FPLC under Denaturing Conditions

.Krimer Alejandro a.krimer at biosidus.com.ar
Thu Mar 4 08:29:41 EST 1999


Dear Michael:

I think you have had some mistake. In general you can elute de proteins with
water in Hydrophobic Interaction Chromatography, like Phenyl-Seph.

Alejandro
> -----Mensaje original-----
> De:	mbarnhart at my-dejanews.com [SMTP:mbarnhart at my-dejanews.com]
> Enviado el:	Wednesday, March 03, 1999 5:00 PM
> Para:	methods at net.bio.net
> Asunto:	Re: FPLC under Denaturing Conditions
> 
> Hi,
> 
> If you want to be certain of denaturation, you should go ahead and use the
> denaturation conditions you would use in other techniques.  One important
> note however, at least one of the FPLC size exclusion columns I used many
> years ago had an affinity for hydrophobic groups.  I never got any protein
> back when I put it on under denaturing conditions but it would come off
> when
> I flushed the column with water.
> 
> Michael
> 
> 
> 
> In article <7lcD2.537$xv.5365831 at WReNphoon2>,
>   ross_turbyfill at wrsmtp-ccmail.army.mil wrote:
> > Does anyone have a protcol for performing FPLC separations (size
> exclusion)
> > under denaturing conditions, i.e., in the presence of guanidine HCl,
> urea,
> > DTT, etc?  Specifically, are the concentrations of the buffers used in
> the
> > chromatography the same concentrations used to denature the proteins
> (i.e.,
> > 8M Urea or 6M GuHCl)?
> >
> > Any advice anyone has to offer would be greatly appreciated!!
> >
> > ross_turbyfill at NOSPAMwrsmtp-ccmail.army.mil
> >
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