I am doing western blots with an RNA binding protein and believe due to the vertical streaks, that I have RNA contamination. I have tried boiling the samples for longer as well. Another protein I have probed for does not have the vertical streaks and is not an RNA binding protein. Could someone recommend a protocol they have used or know of to get rid of RNA contamination in protein preps for western blots?
Hi Anna Schorr , if your protein is prepared by Trizol there should be little-to-no RNA contamination. Have you actually confirmed there is RNA in the sample, or could the streaks just be electrophoretic artifact?
Very simply you could just treat the samples with RNAse A.
As mentioned by Jordan above, use of Trizol for isolation or subsequently treatment with RNase may do the job. However, RNase may not be able to remove the RNA segment bound to an inaccessible binding cavity/site in the protein.
Hello Anna,
what is the problem you experience?
The bands do not fit the expected size?
I do not believe you have an RNA contamination as Jordan R. Yaron poínted out.
Best
Boris
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