Enhanced resolution of phosphoglucomutase (PGM1) by the addition of a separator to ultrathin isoelectric focusing gels

✍ Scribed by Dr. Peter Gill; John G. Sutton

• Publisher: John Wiley and Sons
• Year: 1985
• Tongue: English
• Weight: 523 KB
• Volume: 6
• Category: Article
• ISSN: 0173-0835
• DOI: 10.1002/elps.1150060106

✦ Synopsis

Enhanced separation of phosphoglucomutase (PGM1) 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 PGMl isoenzymes across the whole width ofthe gel. The technique has been shown to be especially useful for separating some ofthe rare PGM, 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 oftemperature on the rate of migration of PGMl isoenzymes has been investigated.