Western blotting - methods to detect two proteins of a similar size? - (Oct/07/2014 )

Hi everyone,

I've run into a bit of a problem with two proteins I'm trying to detect. We have custom polyclonal antibodies against both, raised in rats. I've received test bleeds and detected both proteins; there is no non-specific banding near either band of interest. However, both proteins are very close in size (22 and 23 kDa), and I was hoping to (eventually) detect them on the same blot. I've considered stripping the membrane (starting off with mild stripping reagent), but am unsure if this will work as they're so similar in size. Has anyone had a similar experience/had success in a situation like this?

Hi, have you considered this method ?

http://bitesizebio.com/7846/how-to-transfer-one-sds-page-gel-onto-two-membranes/

Awesome idea, and much better than running two gels instead of one. I'll give it a try soon, thanks!