I am probing prostate cancer cell lines for cytokeratin 5. All 3 showed a band a little larger than anticipated around 60kDA. In one cell line, in addition to the band at 60, there is a heavy signal vertical streak that runs from the well past the band of interest. Has anyone ever seen anything like this? And what could be the cause?
All samples were prepared together, and it is likely not due to boiling/degradation.
Either you loaded to much sample or there is DNA in the sample lysate. The best way to get rid of it is to run the sample several times through a small guage (25G) needle to sheer the DNA
Hi Meghan, I agree with rebecca for the loading problems, it is the first thing, but if you load the same amount for the other samples, and you are sure of your proteins dosage, sometimes vertical streak can appeared for protein precipitation.
Maybe you have this problem only in this sample and the protein precipitates in the staching gel and reaggregated in the running gel. You can try to vortex and spin before after boiling, and it is importat to decrease the voltage in the stacking gel (I have problems with smear and not streak bands, but is quite similar).
Good luck
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