I have actually done this by running a normal PAGE gel without stacking gel and switching the electrodes, which seemed to work swimmingly.
JPK On Thu, Jan 19, 2012 at 9:25 AM, Katherine Sippel <[email protected]> wrote: > 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 > > -- ******************************************* Jacob Pearson Keller Northwestern University Medical Scientist Training Program email: [email protected] *******************************************
