SDS-PAGE precipitation& denaturation - (Nov/08/2006 )

Hello,

I'm going crazy with my SDS-PAGE analysis. If the sample is boiled the protein precipitates and doesn’t enter the gel. Without boiling (or heating) on the other hand, the protein can’t be fully denaturated and therefore it migrates as several bands instead of the expected single band.

How can I solve this problem?

Thanks,

Try to heat the sample to 70°C instead of boiling. It should be also denatured, but perhaps it won't precipitate.

I had the same problem last year (nice small crumbs in the stacking gel), either the SDS buffer was wrong prepared or the SDS concentration was too low. At least, the problem was solved with new SDS-buffer.