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I must have misunderstood you, I loaded 18uL per well (I had 10uL of HeLa nuclear lysate and added 10uL of 2x protein loading buffer to a total volume of 20ul of which I loaded 18ul per well). The wells themselves if loaded carefully can hold 22uL of sample.
Do you recommend I try to load 4uL more?
Asked on Jan 21 2012
Thank you for that information.
I do not think adding 4ul more will increase your signal and since you can only load a maximum of 22ul/well, I would like to send you ab29545 as a free of charge replacement. This HeLa cell lysate is already in sample buffer and is at a higher concentration that the original one sent to you, so you can load more total protein and hopefully see a signal with your 3 antibodies. If this is ok with you, please let me know and I will process the replacement for you.
I look forward to you reply.
Answered on Jan 21 2012