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.
--
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
