Hi Ming,
You can find a protocol for low pH native gel electrophoresis (as well
for high pH and blue native) in the following book:
Gel Electrophoresis of Proteins: A Practical Approach (Practical
Approach Series) by B. D. Hames.
http://www.amazon.com/Gel-Electrophoresis-Proteins-Practical-Approach/dp/0199636400/ref=si3_rdr_bb_product
(Use the amazon online reader and go to page 39).
I hope this helps.
Vangelis
On 26 Aug 2008, at 19:07, Lye, Ming F wrote:
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
