Dear researchers,
When I run SDS-PAGE, sometimes I found that the high molecular weight bands (including bands of the molecular weight marker) disappear from the gel, but low molecular weight bands are visible (see figure below). I've made new running buffer and new component solutions used to make the gel, but this still happen occasionaly.
Has anyone run into the same problem? I would be very appreciated if anyone knows the cause and possible solution. Thanks in advance.
I think somehow your marker and samples have been degraded, making a fresh sample and taking a fresh marker may help you to get rid of this problem.
Samar Gourav Pati Thanks for your suggestion. But I don't think the samples and marker are degraded. Because later I used the same samples and marker to do another run, and it worked well. I found this problem happened randomly even though I've been using the same marker.
Xingchen Chen If I understand you correctly, your problem occurs randomly. To me it looks like that the staining - I guess you have stained your gel with Coomassie blue - is uneven. You need to make sure to use enough Coomassie staining solution to properly cover the entire gel and incubate it on a rocking table/orbital shaker, and avoid laying Kimwipes (or any other wipes) on the gel as this will cause an uneven destaining.
Sabine Strehl Thanks for your suggestions. I did put Kimwipes on the gel to accelerate destaining. I will avoid this in the future and see if the problem occurs again.
@Xingchen chen was your problem resolved? I'm facing same issue. Bands about 66kDa not visible like on your gel. Could you please help me find a solution?
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