Hi Rashmi, In my experience native (even blue native) on proteins around that pI is sketchy at best. The electrophoretic mobility once it gets past the stacking gel goes to crap meaning long electrophoresis times and it needs to be done on a chillable system or in a cold room. If this is a multimer issue I'd suggest trying analytical ultracentrifugation, analytical size exclusion (with the caveat that buffer, temperature and protein shape will affect the output/interpretation), or SAXS first. If native is the only alternative you'll probably get better results changing up the buffer system from traditional tris-glycine or those listed in the blue native protocol keeping in mind that you'll still need to stack the bands.
Good luck, Katherine On Thu, Jan 19, 2012 at 7:03 AM, anita p <[email protected]> wrote: > Hi All, > Has anyone run a native gel for proteins at pI>8 . > I want to pour my own native gel. Do I run a discontinuous page or a > continuous one?? Please help with regards to the buffer system to be used, > and the dye to be used. > With regards > Rashmi >
