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Electrophoresis in one buffer at two pH values

✍ Scribed by Dr. Budin M. Michov


Publisher
John Wiley and Sons
Year
1989
Tongue
English
Weight
371 KB
Volume
10
Category
Article
ISSN
0173-0835

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✦ Synopsis


A theory for discontinuous electrophoresis in a polyacrylamide gel is presented for one buffer at two pH values. It is shown that polyions stack between identical leading and trailing ions, and resolve in a gel of constant polyacrylamide concentration. The theory is illustrated by the separation of serum protein polyions in a Tris-glycinate buffer of pH 8.19 in the well-forming gel, and pH 9.1 6 in the resolving gel. The selected concentrations and electrolyte ionization degrees of Tris and glycine have values at which the serum protein polyions stack between the resolving and electrode buffers, followed by separation in the resolving gel.


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Adequate buffering is of vital importance in capillary electrophoresis. Inadequate buffering may result in severe pH drift and erratic separation behavior. The aim of this work was to study the effects of buffering and buffer replenishment on the repeatability of absolute migration times and on the