Are there any considerations that need to be taken due to the urea or is it a simple transfer? Does anyone know about the interaction between acrylamide and urea?
In my case there was no problem to transfer the target protein and detect it by Western blot. I diluted the 8M urea with loading buffer containing mercaptoethanol 1:2.
Thank you! I've just do it and I've seen that its the same. Have you seen delay in mobility of proteins in the gels with 8M urea? That is, take much longer to run. Thank you!
I run the same speed as samples without urea. But there were some aggregates too big to enter the gel, but a lot of proteins all sizes were seen after staining with ponceau.
you should get rid of the urea coz it will cause distortion of the gel and create fuzzy bands.. u need to wash ur protein at least twice.. and if it dididn't work.. use TCA to get rid of the remaining salts.. Good Luck
I have worked with semi dry electro blotter and did not find any difficulty.
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