Eric,
I second Chun's reply. As the Volumetric flowrate (ml/min) is equal
to the linear flowrate times the area of the column, and the linear
flowrate is fixed for a particular resin (Sigma is terrible at providing
this value for their resins) you basically just need to increase the
diameter of you column to increase the allowable volumetric flowrate.
I typically find that a 2 cm ID column gives a pretty good flowrate
with agarose resins, so much so that I usually use a peristaltic pump to
slow the rate of loading so I have time to drink my morning coffee. As
for washing and elution I usually carry these out at the same flowrate
as loading (including GST resins).
Stephen
--
Stephen Weeks, Ph. D.
Drexel University College of Medicine
Department of Biochemistry and Molecular Biology
Room 10102 New College Building
245 N. 15th St.
Philadelphia, PA 19102
Phone: (+) 215-762-7316
Fax: (+) 215-762-4452
Eric Dollins wrote:
> Dear protein purifiers,
> Off topic question: Is there a general rule for how fast you can load,
> wash and elute from affinity columns, e.g. glutathione agarose? The
> product insert from Sigma says load under gravity flow. For the
> volume of cell lysate I have, gravity loading would take an
> excruciatingly long time. I want to hook up a peristaltic pump to
> speed things along, but don't really have a feel for just how fast one
> can load a column in general (I realize this is also dependent on the
> construct, the buffer, etc). What about the subsequently wash or
> elution?
> Thanks for help
> Eric
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