Colored pI standards and gel isoelectric focusing in strongly acidic pH

Miroslava Štastná, Karel Šlais

Research output: Contribution to journalArticle

Abstract

Colored, low molecular weight pI markers have been developed for isoelectric focusing (IEF) in acidic pH range. Their isoelectric points (pIs) were determined by direct measurement of the pH of the focused bands after completion of IEF on polyacrylamide gels. The practicable suitability of the proposed pI markers as pI standards for IEF was tested by applying gel IEF. The acidic pH gradient was created either by commercial synthetic carrier ampholytes or by mixture of simple buffers consisting of acids (non-ampholytes) and ampholytic buffers. By applying simple acids, it was possible to extend the acidic pH range beyond those achievable with commercial synthetic carrier ampholytes. By using an experimental arrangement without electrode electrolyte reservoirs with electrodes creating the fixed end of the gel, the strongly acidic pH gradient was stable even for prolonged focusing time.

Original languageEnglish (US)
Pages (from-to)65-72
Number of pages8
JournalAnalytical and Bioanalytical Chemistry
Volume382
Issue number1
DOIs
StatePublished - May 2005
Externally publishedYes

Keywords

  • Acidic pH
  • Colored pI markers
  • Gel isoelectric focusing

ASJC Scopus subject areas

  • Analytical Chemistry
  • Clinical Biochemistry

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