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
<[log in to unmask]> 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 <[log in to unmask]> 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: [log in to unmask]
*******************************************
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