Size exclusion chromatography for 23 and 20.5KDa? - protein purification (Sep/19/2007 )

Hi, I have two proteins that are 23KDa and 20.5KDa in size. Is it possible to seperate them using a Superdex 200 10/300 GL column in FPLC? Anyone has experiences with it, or columns to recommend? Thanks guys!

-Tummybear-

you probably won't see any significant separation with superdex 200. you may be able to get a good enough separation with superdex 75 (if you keep your injection volume low and not too much protein).

-mdfenko-

Agree with Mdfenko. Superdex 75 will be better ( because Kav for this Mw range is in optimal range ( 0.4-0.5) or as alternative may be Superose 12, but in general it will be better to change method of separation ( by charge IEC chromatography) or by hydrofobicity ( HIC chromatography). I think it will be
a short way to separation of closed Mw proteins.

-circlepoint-

Hey guys.. thx for the input - appreciate it.

-Tummybear-