Column Choice

Leemor Joshua-Tor leemor at CSHL.ORG
Mon Sep 16 10:04:50 EST 1996

Dima Klechin wrote:

>In article <51i1cl$1r6 at>, lhom at
>(Louis Hom) wrote:
>#I need to run a sizing column to see if my ca. 30kD protein is associated
>#with other proteins.  Can folks recommend places to look where I can find
>#out what the best matrix and respective column sizes would be?  (I might be
>#running this by gravity, maaaybe by FPLC, depending on things.)
>Well, if FPLC, then only Superose 12 makes choice.
>If by gravity, then the common place to start with 30 K is
>Sephacryl 200 (depending on the need of resolution, 100 and 300 might
>be better). Sepharose 6B is too porous while Sephadexes 150 and 200 are
>pain in the neck.
>As far as column size goes, it depends on the volume of your sample.
>Generally, to achieve reasonable resolution, loading volume should be < 3%
>of column bed.  Commonly used ratios  of diameter to length are 1:10 -
>1:50 depending on application.
>- Dima

Actually, Pharmacia has more choices now, like superdex, which works
pretty well for us. I suggest giving their tech support a call. Some
of them are pretty helpful.

  Leemor Joshua-Tor, Ph.D.
  Assistant Investigator
  Cold Spring Harbor Laboratory           Tel. (516) 367 8821
  1 Bungtown Road                         Fax  (516) 367 8873
  Cold Spring Harbor, NY 11724            e-mail: leemor at

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