Re: [ccp4bb] Superdex 200 PC columns

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From: "Y {- dot -} -F {- dot -} Li" yongfuli {- at -} GMAIL {- dot -} COM
Date: 2007-05-12

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Subject: Re: S-tagged protein purification
From: artem {- at -} XTALS {- dot -} ORG artem {- at -} XTALS {- dot -} ORG
Date: 2007-05-12

Subject: Re: Superdex 200 PC columns
From: artem {- at -} XTALS {- dot -} ORG artem {- at -} XTALS {- dot -} ORG
Date: 2007-05-12

It greatly depends on the state of your instrument - how well you took
care of the optical elements, etc. It also depends on the initial
concentration of the sample (and therefore the peak volume at detection
time) and on the optical activity of the sample and the buffer. If the
stars align and everything works, you should be able to easily detect 5 ug
of protein in a *reasonable* peak volume. This would require a true
injection of perhaps 15 ul assuming a dilution down to about 50 ul in the
peak. Oviously if you have multiple smeary peaks than you won't get any
signal at all :)

Artem

> 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.
>

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CCP4bb <-- 2007 <-- May 2007 <-- 12 May 2007

Previous message:
Subject: S-tagged protein purification
From: "Y {- dot -} -F {- dot -} Li" yongfuli {- at -} GMAIL {- dot -} COM
Date: 2007-05-12

Next message:
Subject: Re: S-tagged protein purification
From: artem {- at -} XTALS {- dot -} ORG artem {- at -} XTALS {- dot -} ORG
Date: 2007-05-12