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I'm currently trying to detect a phosphorylated-myosin light chain protein that runs around 18 kDa. I just processed a couple of blots and didn't really see much protein, leading me to believe that there might have been a problem in transfer.
I'm using an 18% polyacrylamide gel and transferred like I normally do with 10% gels (100 Volts x 60 minutes at 4 degrees), transferring to nitrocellulose. I'm thinking this is too short of a time for such a high percentage gel. Just wondering if people who had experience with high percentage gels/low MW proteins had any input regarding their protocol vs. what I'm doing. Thanks!
