IUBio

FPLC under Denaturing Conditions

.Krimer Alejandro a.krimer at biosidus.com.ar
Thu Mar 4 08:13:23 EST 1999


The best way is to equilibrate and run the separation with the same
concentration of guanidine or urea.
I have worked with several proteins without any problems, 
Hope this helps.

						Alejandro
> -----Mensaje original-----
> De:	ross_turbyfill at wrsmtp-ccmail.army.mil
> [SMTP:ross_turbyfill at wrsmtp-ccmail.army.mil]
> Enviado el:	Wednesday, March 03, 1999 10:40 AM
> Para:	methods at net.bio.net
> Asunto:	FPLC under Denaturing Conditions
> 
> 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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