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Hi,
You're right, there are two cell path sizes in AKTA systems: 2 mm and 10
mm. I don't know what path had the SMART. You definitely need the second
one if you want to work with low protein concentrations.
Miguel
En/na Juergen Bosch ha escrit:
> 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
>
> [EMAIL PROTECTED] 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.
>>>
>>>
>>
>>
>>
>
>
- --
Miguel Ortiz Lombardía
Centro de Investigaciones Oncológicas
C/ Melchor Fernández Almagro, 3
28029 Madrid, Spain
Tel. +34 912 246 900
Fax. +34 912 246 976
e-mail: [EMAIL PROTECTED]
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Et ainsi ne pouvant faire que ce qui est juste fût fort, on a fait que
ce qui est fort fût juste.
Blaise Pascal, Pensées
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