Activity staining of isocitrate lyase after electrophoresis on either native or sodium dodecyl sulfate polyacrylamide gels

Wen Chi Hou, Hsien Jung Chen, Yaw Huei Lin, Yen Chou Chen, Ling-Ling Yang, Mei Hsien Lee

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Abstract

Isocitrate was cleaved into succinate and glyoxylate by isocitrate lyase (ICL) in the glyoxylate cycle. We used lactate dehydrogenase as an ancillary enzyme to further metabolize the glyoxylate to glycolate in the presence of NADH. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 2,6-dichlorophenol-indolphenol (DCPIP) were used in the coupling reactions for detecting ICL activity after electrophoresis on either native or sodium dodecyl sulfate (SDS) polyacrylamide gels. This fast and sensitive method can be used in the process of ICL enzyme purification and characterization.

Original languageEnglish
Pages (from-to)2653-2655
Number of pages3
JournalElectrophoresis
Volume22
Issue number13
DOIs
Publication statusPublished - 2001

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Keywords

  • Activity staining polyacrylamide gel electrophoresis
  • Isocitrate lyase
  • Native sulfate-polyacrylamide gel electrophoresis
  • Sodium dodecyl

ASJC Scopus subject areas

  • Clinical Biochemistry

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