Quick navigation:        Home   |    Site Map   ||    References   |    Biography   ||    Copyright   |    Other copyright   |    Contact us   |   
Protein structure
 

Re: [ccp4bb] Off topic: General rule for maximum flow rate for affinity column?
 

Basic tutorials: 		 
 

CCP4bb navigation

CCP4bb <-- 2008 <-- February 2008 <-- 28 February 2008
Previous message:
Subject: O-glycosylation in ccp4/refmac
From: Rebecca Phillips rephilli {- at -} CHEM {- dot -} UCSD {- dot -} EDU
Date: 2008-02-28 		Next message:
Subject: Re: Off topic: General rule for maximum flow rate for affinity column?
From: Artem Evdokimov artem {- at -} XTALS {- dot -} ORG
Date: 2008-02-28


Subject: Re: Off topic: General rule for maximum flow rate for affinity column?
From: Chun Luo chun {- at -} ACCELAGEN {- dot -} COM
Date: 2008-02-28
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
chun@accelagen.com
www.accelagen.com


-----Original Message-----
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] 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, d.e.dollins@gmail.com

CCP4bb navigation

CCP4bb <-- 2008 <-- February 2008 <-- 28 February 2008
Previous message:
Subject: O-glycosylation in ccp4/refmac
From: Rebecca Phillips rephilli {- at -} CHEM {- dot -} UCSD {- dot -} EDU
Date: 2008-02-28 		Next message:
Subject: Re: Off topic: General rule for maximum flow rate for affinity column?
From: Artem Evdokimov artem {- at -} XTALS {- dot -} ORG
Date: 2008-02-28


ProteinCrystallography.org: Copyright 2006-2008 by Quid United Ltd