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Native polyacrylamide electrophoresis in the presence of Ponceau Red to study oligomeric states of protein complexes
Author(s) -
Dráb Tomáš,
Kračmerová Jana,
Tichá Ivana,
Hanzlíková Eva,
Tichá Marie,
Liberda Jiří
Publication year - 2011
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201000869
Subject(s) - chemistry , polyacrylamide , polyacrylamide gel electrophoresis , chromatography , gel electrophoresis of proteins , polymer chemistry , biochemistry , enzyme
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.