FPLC under Denaturing Conditions

mbarnhart at my-dejanews.com mbarnhart at my-dejanews.com
Wed Mar 3 15:00:21 EST 1999


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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