Acid native-PAGE buffer(s)
hiroki at limerick.cbs.umn.edu
Sat Jul 12 02:19:51 EST 1997
The author you probably want to look up is
Jovin, from way back when. Analytical Biochem I think.
If it's really 8.8-9, you'll probably get decent
resolution even at pH 7 or so.
I've used something like 25mM His, 25mM Mops throughout
(or was it MES?) at about pH 6.2 or so.
Running buffer same as in the gel.
Didn't really bother trying to make a
real stacking gel, I just cut back on the acrylamide,
and got good resolution, so stopped at that point.
75% Glycerol 25% of the buffer, and a smidgen of methylgreen
for the loading dye.
I know it's not a discontinuous acid system, the only one I vaguely
remember is supposed to form "dumbell" shaped bands.
steve (dunnsm at bbsrc.ac.uk) wrote:
: Hello All,
: I'm planning on performing a native-PAGE expt. on a couple of proteins
: with fairly high predicted pI values (approx. 8.8-9.0) and so I'm looking
: for an acid buffer system that operates at around pH 5-6 (I'd rather not
: go more acid than this). Does anybody have tried and tested recipes for
: a discontinuos system in this pH range?
: Many thanks for any info/advice.
: dunnsm at bbsrc.ac.uk
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