FPLC with Denaturing Conditions
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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