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Hi Robert,
I have run those PC 2.3/30 with ~10 ug of protein (of course this
depends on the protein) and still get a useful signal at 280 nm. If you
don't have anything interfering at ~215 nm and your HPLC system can read
at that wavelength, you can even do with less than that. They are very
useful columns, for example to check for interactions, but they need to
be handled with _extra_ care. Your HPLC system as well needs to be very
stable and _really_ clean, especially the photometer cell. You should
use loops of 10-25 ul, though your sample may need to be bigger to avoid
getting "lost" in dead spaces.
Good luck,
Miguel
En/na Robert Cleverley ha escrit:
> 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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