I am running some western blots currently and need to detect 2 proteins with relatively different molecular weight. one is 63KDa and the other is 262KDa. I am looking for ways to modify my protocol with out having to run 2 separate gels. Does anyone have some insights to how I may achieve this. I already have an established protocol to the detect the lower MW protein but do not know how to incorporate detecting the higher MW protein into the same protocol..
Thank you!
Can you cut the membrane near below the standard at 100 KDa, after transferred it? Then incubate each piece of membrane separately.
But you can't cut the membrane then continue to transfer, because when you cut you'll never get it right the membrane in the same place as before. Do you know any protein that you could use as endogenous with a higher molecular weight above 80 kDa?
Other solution is to use the WES system, but for this you need to know someone that have this machine.
I see what you mean it wilbe be hard to get the membrane back to the exact position. how about cutting the gel and transferring separately?
Ok, maybe in this manner it will work. But, you will still have the same problem with the running, I mean, if you run the gel for more than 1 hour and half, you probably lose your low MW protein anyway. Good luck.
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