Has anybody seen the jiggling of FPLC trace like this? I have cleaned the flow cell which was hinted to be the cause of this with 0.5 M NaOH followed by 500 ml wash with MilliQ water. Still, I am seeing the same.
Dear Niyati Jain
Most of the noise is within the range of 10 mAU, so I would not worry much about that. What comes to my mind is that possible reason for such signal is just electronic noise (detector) or not sufficiently degassed buffers.
Thanks, Tomasz for your answer but due to this sometimes the UV detector is not giving the signal for the sample and I just see the noise when the sample is passed through the analytical column.
Dear Niyati,
Does the pressure fluctuate as well? this could be a sign of air bubbles in your system like Tomasz Fraczyk mentioned. You could try washing your system with DDW, then pure methanol (NOT through column, system wash only), then DDW. Check the manual first, though (works with AKTA go, it seems you have an older AKTA model).
Alternatively, your UV-lamp may be dying. They can be noisy near the end. Try purifying a common protein like BSA to see if you receive a signal.
Thanks, Roy the flow cell looked like the culprit and for now, the trace looks ok washing it with 1M NaOH 30 ml at 1ml/min with flow restrictor in place followed by 60 ml of water at 4-5 ml/min flow rate helped to remove the air bubble.
I think you should check the lamp life, maybe it is dying. It could also be a problem with the pressure or the composition of your buffer. Do you also have this problem when you are running with water ?
Hi, Isabel yes I have a problem with water as well it seemed to work fine for a few days but again the problem is coming back.
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