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Activity staining of isocitrate lyase after electrophoresis on either native or sodium dodecyl sulfate polyacrylamide gels
Author(s) -
Hou WenChi,
Chen HsienJung,
Lin YawHuei,
Chen YenChou,
Yang LingLing,
Lee MeiHsien
Publication year - 2001
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200108)22:13<2653::aid-elps2653>3.0.co;2-o
Subject(s) - glyoxylate cycle , isocitrate lyase , sodium dodecyl sulfate , chemistry , polyacrylamide gel electrophoresis , gel electrophoresis , biochemistry , chromatography , isocitrate dehydrogenase , polyacrylamide , bromide , enzyme , staining , sodium , biology , organic chemistry , polymer chemistry , genetics
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.