Native/ Nondenaturing PAGE - (Jul/13/2007 )

Can anyone suggest a protocol for running a Native/Nondenaturing PAGE? I am having trouble finding a suitable sample buffer, and I'd like to transfer the gel to a nitrocellulose membrane but haven't had any luck.
Thanks!

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there is no common protocol as f.i. in SDS-PAGE as in native PAGE the pI of your protein(s) of interest decides which buffer you need...

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So the pI of my protein of interest is 8.9, and I have tried transferring my gel to nitrocellulose in 0.7% acetic acid (with the electrodes reversed) for 1 hr at 100V, but that was unsuccessful. Any ideas?

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see this link for the formulation of an acidic page method:

acidic page

you do not need any sample buffer, just add glycerol and tracking dye to your sample and run.

as for transfer, you could try the same electrode buffer as with the gel.

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