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Enhanced resolution of phosphoglucomutase (PGM 1 ) by the addition of a separator to ultrathin isoelectric focusing gels
Author(s) -
Gill Peter,
Sutton John G.
Publication year - 1985
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150060106
Subject(s) - phosphoglucomutase , isoelectric focusing , electrolyte , chemistry , histidine , chromatography , hydroxide , immobilized ph gradient , isoelectric point , sodium hydroxide , analytical chemistry (journal) , inorganic chemistry , biochemistry , amino acid , enzyme , organic chemistry , electrode
Enhanced separation of phosphoglucomutase (PGM 1 ) isoenzymes has been achieved by addition of the separator N‐(2‐Hydroxyethyl)piperazine‐N′‐3‐propanesulphonic acid (EPPS) to pH 5–7 Ampholines in ultrathin and 1 mm isoelectric focusing gels. The resultant pH gradient ranges between pH 5.6 to pH 5.9 and effectively spreads PGM 1 isoenzymes across the whole width of the gel. The technique has been shown to be especially useful for separating some of the rare PGM 1 variants. The use of different electrolyte combinations has also been investigated and it has been shown that strong electrolytes, e. g. sodium hydroxide and orthophosphoric acid, result in a small but statistically significant increase in the gradient range compared with weaker electrolytes such as threonine/histidine. In addition, the effect of temperature on the rate of migration of PGM 1 isoenzymes has been investigated.