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Some bands transferred, some not..? Why???

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#1 bongiwoman



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Posted 13 August 2015 - 09:36 PM

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!



#2 mdfenko


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Posted 14 August 2015 - 03:21 AM

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)

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