Native PAGE and WB - (Apr/06/2007 )
Hi, I am dealing with a mammalian neuronal membrane transport protein, and would like to do a native PAGE/WB analysis to determine the presence of multimeric versus monomeric forms in tissue and cell culture. Does anyone have any recommendations for lysate buffer and solubilization protocols I can use without denaturants? Also, I read somewhere that the transfer buffer doesn't require methanol if you're doing native PAGE... is that really so?
Thanks for any and all help,
yes. one function of methanol in the transfer is that it helps strip the sds off of the protein to enhance binding (this may be more important for nitrocellulose).