FPLC with Denaturing Conditions

VVS vvs at sun.ipr.serpukhov.su
Thu Mar 4 03:42:56 EST 1999


No problem! You can see in the manual for your column all parameters
  of it. I sink what could be better use buffer contains about 100 -
  200 mM KCl or NaCl. The volume for equilibration of your column you
  will know after reading of the manual.
  Caution: do not use MAXIMAL flow rate if you use buffer with high
  concentration  of urea or GuHCl!

>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)?





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