Some bands transferred, some not..? Why??? - (Aug/14/2015 )

Hello researchers,

I did a Western with several lysates of different cell lines, always treated with DMSO or a drug (always the same).

After Ponceau staining, I see that some cell lines did not get transferred - either the high molecular proteins or even any bands. I treated all cell pellets the same way - used RIPA buffer to lyse them and added the same sample buffer, so I really don't have a clue what happened...? It does not look like bubbles or anything because it is very restricted to the two lanes of the same cell lines.

Thanks a lot for any advice!

are you sure that you loaded protein in those lanes (do you have a stained gel or have you stained the gel post-transfer)?

there may be too little to see with ponceau. have you tried immunostaining?

(pictures would help with diagnosis)