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 > > > > > > > >
