Hi Chiara,
We also have an Avant at room temperature. My recommendation would be to
forgo using a water bath. Instead you can achieve a prep near enough to 4C
by doing 2 things: Firstly, keep your columns stored in the cold room or a
fridge to pre-chill them before use. Secondly, make your buffers up with
chilled water and then either put them in an ice bucket or you could get a
bottle jacket (much like a wine bottle cooler) and use that. If you want to
make extra sure of a low temperature, you could keep the ice frozen longer
using some cold packs from the freezer. GE have tested this setup with some
thermocouples and find the temperature drop across the column goes from 6C
at the top to 7C at the bottom even with a 16/600 column, so it should
definitely be fine for a shorter 10/300.
Having said all that, our local Äkta specialist from GE assures us that,
except for pathological cases, room temperature preps are generally OK,
especially if your protein is immobilised on a column - proteins are pretty
stable under those conditions. And, of course, the fraction collector can
be chilled to 6C, so as soon as your samples come off the column they will
be chilled.
--
Roger B. Dodd, PhD
Cambridge Institute for Medical Research
University of Cambridge School of Clinical Medicine
Wellcome Trust/MRC Building
Cambridge Biomedical Campus
Hills Road
Cambridge
CB2 0XY
On Tue Feb 03 2015 at 10:15:26 AM Chiara Rapisarda
chiara.rapisa...@pasteur.fr wrote:
Hello,
we have an AKTA Avant that we keep at room temperature. We would like to
be able to use it for preps at 4 degrees, so we would like to keep the FPLC
columns (type 10/30 from GE) in the cold. We were thinking of buying a
circulating water bath and connect it somehow to the columns through a
jacket. Has anybody had any experience with that? We looked also into
buying the hot pocket from Fisher Scientific but it is a bit out of our
budget.
Any suggestions are very welcome.
Chiara
POSTDOC
Structural Biology of
Bacterial secretion
Department of Structural Biology and Chemistry
INSTITUT PASTEUR
25 rue du Docteur Roux
75015 Paris FRANCE
*Dr. Chiara Rapisarda*
*Tel : +33 (0)1.40.61.35.31*
*Fax :**+33 (0)1.45.68.88.36*
*Email: chiara.rapisarda*@pasteur.fr
