After removing a plug from a broken Ni column from our FPLC UV sensor, all elution profiles of subsequent chromatography runs (both Ni column and SEC purification) appear jagged and distorted. The two pictures attached below show a "pristine" elution profile (top) for one of our proteins and a "jagged" one (bottom) showing a more distorted profile for that same protein after the incident with our broken Ni column. We believe that there might be an issue with the sensor.
Has anyone else experienced something like this? If so, is this an easy fix?
In case this may be relevant, I should add that we are running denaturing purification with 4M guanidinium chloride in our buffers.
What is the scale of UV? Is one higher than the other?
I would try check for air bubble. You may add restrictor (e.g. original, or make one from long and thin tube), to push out air bubbles.
- Badges
- Science topic