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Re: [ccp4bb] Superdex 200 PC columns
 

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CCP4bb <-- 2007 <-- May 2007 <-- 12 May 2007
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Subject: Re: Superdex 200 PC columns
From: mlamers {- at -} BERKELEY {- dot -} EDU mlamers {- at -} BERKELEY {- dot -} EDU
Date: 2007-05-12 		Next message:
Subject: Re: Superdex 200 PC columns
From: Miguel Ortiz-Lombardia molatwork {- at -} YAHOO {- dot -} ES
Date: 2007-05-12


Subject: Re: Superdex 200 PC columns
From: Juergen Bosch jbosch {- at -} U {- dot -} WASHINGTON {- dot -} EDU
Date: 2007-05-12
Hi all,

Isn't the flow cell volume the limiting factor for your detection limit
? I believe for the "old" Aktas (~5-8 years old) there were two cell
sizes a laerger one and a smaller one, and I don't think it is the same
size as in the SMART system. But I might be wrong.

Jürgen

mlamers@BERKELEY.EDU wrote:

>Hi Robert,
>
>In my experience, there is no problem running a SMART column on an AKTA
>FPLC or Explorer. I have been using the same S75, S200, and Superose6
>columns on all three systems depending on which one is available that day.
>I typically load 50 ul sample, between 10-0.01 mg/ml. The peak width I get
>is ~150ul, with 70-80% of the original concentration at the height of the
>peak.
>On the FPLC/Explorer I replace the standard tubing to shorter and more
>narrow tubing and bypass the conductivity meter to reduce the amount of
>dead volume.
>Finally, for the very low protein concentrations, you're better of using
>the Explorer, which allows you to measure the absorption at 220-230nm.
>
>
>Meindert
>
>
>
>
>>Sorry for perhaps off-topic question but I am writing to ask if anyone has
>>experience in using Amersham Superdex 200 PC 3.2/30 columns in conjuction
>>with
>>the Precision Column holder on a standard (not SMART system) AKTA FPLC.
>>At the
>>moment using a Superdex 200 10/300 column on a standard AKTA I can get a
>>reasonable signal from down to about 20 ug protein - I am wondering how
>>much
>>less protein I will be able to use with the narrower Superdex 200 PC
>>3.2/30
>>column when I hook it up to the same FPLC via the Precision Column holder.
>>
>>
>>
>
>
>


--
Jürgen Bosch
University of Washington
Dept. of Biochemistry, K-426
1705 NE Pacific Street
Seattle, WA 98195
Box 357742
Phone: +1-206-616-4510
FAX: +1-206-685-7002

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CCP4bb <-- 2007 <-- May 2007 <-- 12 May 2007
Previous message:
Subject: Re: Superdex 200 PC columns
From: mlamers {- at -} BERKELEY {- dot -} EDU mlamers {- at -} BERKELEY {- dot -} EDU
Date: 2007-05-12 		Next message:
Subject: Re: Superdex 200 PC columns
From: Miguel Ortiz-Lombardia molatwork {- at -} YAHOO {- dot -} ES
Date: 2007-05-12


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