FPLC is behaving strangely - Slow running (Aug/21/2009 )

We'll after cleaning it with NaOH wash protocol, I equilibrated with my buffer and ran a prep.
And I increased the pressure alarm... to 0.3 MPa (under supervision).

The prep worked out fine, and I got a crisp peak. No annoying overpressures every 10 mL.
You could still see tiny pressure spikes, but these instantaneously return to normal baseline pressures. Must be something quirky with the machine or resin.

Finally. It works. I'm very happy
Thanks for the advice everyone.

LB

* Always call up the manufacturer and work under supervision before doing. I can't accept any liability if you follow this advice, damage results or guarantee that it is correct.*

Luria Bertani on Sep 5 2009, 05:25 AM said:

Finally. It works. I'm very happy

I don't doubt that your column had some blockage, but if you are using an Akta FPLC and the spikes are about every 10ml you also may want to turn pulse compensation off. This is especially true if you are running columns with max pressure of 0.5MPa or less.