Tips on Western Transfers (iBlot2 or old fashion way?) and multiplexing

Does anybody have any insight/protocol optimizations on western transfers ( I have an iBlot2 but can do it the old fashion way too). When staining the gel post-transfer it looks that 80-90% of the protein remains on the gel with minimal to no protein is on the membrane. Looking to identify proteins in the 30-60kDa range and loading 40-50ug of protein on the gel.


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Pavitra Viswanathabout 1 year ago

With the old fashioned wet transfer, using 100V for 90 min on ice/cold has been giving me great transfers after loading 50ug of protein, especially the 30kd ones and the big ones over 150kd. That might hopefully work for you?

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