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Re: [ccp4bb] Off topic: General rule for maximum flow rate for affinity column? |
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CCP4bb navigationCCP4bb <-- 2008 <-- February 2008 <-- 28 February 2008Subject: Re: Off topic: General rule for maximum flow rate for affinity column? From: Stephen Weeks stephen {- dot -} weeks {- at -} VERIZON {- dot -} NET Date: 2008-02-28 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 > > > CCP4bb navigationCCP4bb <-- 2008 <-- February 2008 <-- 28 February 2008 |
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