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Native polyacrylamide electrophoresis in the presence of Ponceau Red to study oligomeric states of protein complexes.

Journal of separation science (2011-06-18)
Tomáš Dráb, Jana Kračmerová, Ivana Tichá, Eva Hanzlíková, Marie Tichá, Jiří Liberda
ABSTRACT

Native polyacrylamide electrophoresis in the presence of two reversible protein anionic stains (Ponceau S and Ponceau 2R) was used to study the oligomeric states of soluble proteins. A mild binding of the used protein stains to nondissociated protein oligomers imposed a charge shift on the proteins resulting into separation of protein species according to their size under physiological conditions. Adsorbed stains could be easily removed after electrophoresis by washing of polyacrylamide gel with buffer and protein complexes could be visualized either by the detection of their enzyme activity or by using a nonspecific protein stain. The specific detection of enzyme activity of glycosidases, lactate dehydrogenase, or phosphatases was shown as an example.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ponceau S solution, BioReagent, suitable for electrophoresis, 0.1 % (w/v) in 5% acetic acid
Sigma-Aldrich
Ponceau S, BioReagent, suitable for electrophoresis
Sigma-Aldrich
Ponceau S, Dye content 75 %

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