You could try Coomassie Blue Native gel. It's a very neat technique and it worked for me on a couple of occasions. In one unfortunate case, it resulted in dissociation of a heterotetrameric complex, though.
Artem > Dear CCP4 community, > > Sorry for the off-topic subject, but I would really appreciate some > suggestions and/or protocols relating to native gel electrophoresis of > basic proteins. I have used a general acidic PAGE protocol for my protein, > which has a PI of 9.5. Briefly, the protein was loaded onto a native gel > (I have tried both the pre-made Biorad gels (7.5% and a gradient gel: > 4-15%) and freshly prepared Tris native gels adjusted to pH 6.8) and run > in a 1X acetic acid/b-alanine pH 4.5 running buffer. The electrodes were > reversed and the gel run on ice for ~ 2hrs at 100V. In all cases, the > native protein was unable to enter the gel. Some protein samples incubated > with heavy atoms were able to enter the gel (possibly indicating binding) > but these samples too had problems entering the gel as the bands were at > or just a little bit below the edge of the well. Any suggestions and > comments would be most welcome! > > Thank you so much in advance for your help, > Sincerely, > Ming Lye >
