65 and 64 separation
hroychow at NMSU.EDU
Mon Aug 8 12:09:50 EST 1994
On 5 Aug 1994, jinsong sheng wrote:
> Dear Netters:
> Any suggestions to separate a 65 and 64 kd proteins on SDS -PAGE? Like
> percentage of gels, gradient or buffer systems?
> Many thanks in advance.
> Jinsong Sheng.
The % of the gel can be varied and the separation tried out. I would try
three different gradients: 10-15%, 10-12% and 8-12%. However, since the
separation is often dependent on the type of protein (state of conjugation
etc.), one can also obtain reasonable separation by just an isocratic gel.
Again, the % T has to be chosen by trial-and-error.
If this simple step does not help then one can manipulate the %C (ie.
instead of a regular 4 or 4.5%C).
Tricine buffer system (instead of glycine), which also employs a higher
%C, has been known to separate smaller polypeptides (below 12 kDa) rather
efficiently. I am not sure if it will work for moderately higher mol. wt.
polypeptides. Worth a try.
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