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

doi:10.1002/1522-2683(200108)22:13<2653::AID-ELPS2653>3.0.CO;2-O

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

Research output: Contribution to journal › Article › peer-review

6 Citations (Scopus)

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 language	English
Pages (from-to)	2653-2655
Number of pages	3
Journal	Electrophoresis
Volume	22
Issue number	13
DOIs	https://doi.org/10.1002/1522-2683(200108)22:13<2653::AID-ELPS2653>3.0.CO;2-O
Publication status	Published - 2001

Keywords

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

ASJC Scopus subject areas

Analytical Chemistry
Biochemistry
Clinical Biochemistry

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10.1002/1522-2683(200108)22:13<2653::AID-ELPS2653>3.0.CO;2-O

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

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.",

keywords = "Activity staining polyacrylamide gel electrophoresis, Isocitrate lyase, Native sulfate-polyacrylamide gel electrophoresis, Sodium dodecyl",

author = "Hou, {Wen Chi} and Chen, {Hsien Jung} and Lin, {Yaw Huei} and Chen, {Yen Chou} and Ling-Ling Yang and Lee, {Mei Hsien}",

year = "2001",

doi = "10.1002/1522-2683(200108)22:13<2653::AID-ELPS2653>3.0.CO;2-O",

language = "English",

volume = "22",

pages = "2653--2655",

journal = "Electrophoresis",

issn = "0173-0835",

publisher = "John Wiley and Sons Inc.",

number = "13",

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TY - JOUR

T1 - Activity staining of isocitrate lyase after electrophoresis on either native or sodium dodecyl sulfate polyacrylamide gels

AU - Hou, Wen Chi

AU - Chen, Hsien Jung

AU - Lin, Yaw Huei

AU - Chen, Yen Chou

AU - Yang, Ling-Ling

AU - Lee, Mei Hsien

PY - 2001

Y1 - 2001

N2 - 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.

AB - 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.

KW - Activity staining polyacrylamide gel electrophoresis

KW - Isocitrate lyase

KW - Native sulfate-polyacrylamide gel electrophoresis

KW - Sodium dodecyl

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U2 - 10.1002/1522-2683(200108)22:13<2653::AID-ELPS2653>3.0.CO;2-O

DO - 10.1002/1522-2683(200108)22:13<2653::AID-ELPS2653>3.0.CO;2-O

M3 - Article

C2 - 11545388

AN - SCOPUS:0034845111

SN - 0173-0835

VL - 22

SP - 2653

EP - 2655

JO - Electrophoresis

JF - Electrophoresis

IS - 13

ER -

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

Abstract

Keywords

ASJC Scopus subject areas

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