Hi Eric, Check the max pressure of the resin. Some tell you the flow-rate in cm/hr also. You can calculate that to ml/min with your column dimension. Most resins can stand 5 ml/min flow-rate in an ID2.5cm column.
However, most people will recommend a slower flow-rate for glutathione agarose, claiming slow on rate. That's a myth. In our hands, glutathione resins binds as fast as Ni resins. Chun Chun Luo, Ph.D. The Protein Expert Accelagen, Inc. 11585 Sorrento Valley Road, Suite 107 San Diego, CA 92121 TeL: 858-350-8085 ext 111 Fax: 858-350-8001 [EMAIL PROTECTED] www.accelagen.com -----Original Message----- From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of Eric Dollins Sent: Thursday, February 28, 2008 2:26 PM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] Off topic: General rule for maximum flow rate for affinity column? 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 -- D. Eric Dollins, Ph.D. C266 LSRC, Research Dr. Duke University Medical Center Durham, NC 27710 (919) 681-1668, [EMAIL PROTECTED]
