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
