Limitations of Western Blotting | Homeworkmaven.com

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Why am I able to visualize my protein ladders but unable to visualize GAPDH on my western blot?

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What voltage did you use for the transfer? GAPDH is a small protein, and does not necessarily need to be transferred overnight, it is possible that you over transferred? I usually do a wet transfer and transfer for 80 min at 80 volts. You can also stain the membrane (before incubation with block) with Ponceau S stain to see if it is the transfer.

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