Activity staining of isocitrate lyase after electrophoresis on either native or sodium dodecyl sulfate polyacrylamide gels
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) a...
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| Veröffentlicht in: | Electrophoresis 2001-08, Vol.22 (13), p.2653-2655 |
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| creator | Hou, Wen-Chi Chen, Hsien-Jung Lin, Yaw-Huei Chen, Yen-Chou Yang, Ling-Ling Lee, Mei-Hsien |
| description | 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. |
| doi_str_mv | 10.1002/1522-2683(200108)22:13<2653::AID-ELPS2653>3.0.CO;2-O |
| format | Article |
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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. 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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.</description><subject>Acrylic Resins</subject><subject>Activity staining polyacrylamide gel electrophoresis</subject><subject>Electrophoresis, Polyacrylamide Gel - methods</subject><subject>Isocitrate lyase</subject><subject>Isocitrate Lyase - analysis</subject><subject>Native sulfate-polyacrylamide gel electrophoresis</subject><subject>Protein Denaturation</subject><subject>Sodium dodecyl</subject><subject>Sodium Dodecyl Sulfate</subject><subject>Staining and Labeling - methods</subject><issn>0173-0835</issn><issn>1522-2683</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqVkU2P0zAQhiMEYsvCX0A-ITikjO04H2WFVJWlLKq2lVgJOI1cZ7JrSOtiJ0D49bhqKScOnKzXHj-vNE-SXHAYcwDxkishUpGX8rkA4FC-EGLC5YXIlZxMpldv0svF6sM-vZZjGM-Wr0S6vJeMTt_uJyPghUyhlOoseRTCFwDIqix7mJxxrjIly3KU_Jqazn633cBCp-3Wbm-Za5gNztjO645YO-hATDcdeUYtmc673Z3zFGxgbsvIdnfxZasjhZjzLLja9htWu5rM0LLQt80es3MRZPzQ6o2tid1SGx4nDxrdBnpyPM-Tm7eXN7N36WI5v5pNF6nJIJNpVcpSNWDWNQnOTU1a8EZLMqXQCpQs8hwaDgqg0mtRqExUa1OpklfFmgPJ8-TZAbvz7ltPocONDYbaVm_J9QGLuAyuci5P_ca7EDw1uPN2o_2AHHCvBPe7xf1u8aAEY-QxRAmIUQn-UYISAWdLFLiM2KfH_n69ofov9OggDnw-DPywLQ3_VfqPztNdZKcHtg0d_Tyxtf-KeSELhR-v57harMQ1fz_HT_I34XO3hA</recordid><startdate>200108</startdate><enddate>200108</enddate><creator>Hou, Wen-Chi</creator><creator>Chen, Hsien-Jung</creator><creator>Lin, Yaw-Huei</creator><creator>Chen, Yen-Chou</creator><creator>Yang, Ling-Ling</creator><creator>Lee, Mei-Hsien</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>200108</creationdate><title>Activity staining of isocitrate lyase after electrophoresis on either native or sodium dodecyl sulfate polyacrylamide gels</title><author>Hou, Wen-Chi ; Chen, Hsien-Jung ; Lin, Yaw-Huei ; Chen, Yen-Chou ; Yang, Ling-Ling ; Lee, Mei-Hsien</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4043-98385f0cbde211cdea21fa3ec82a50537660f105009ab275429bc958197b10e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>Acrylic Resins</topic><topic>Activity staining polyacrylamide gel electrophoresis</topic><topic>Electrophoresis, Polyacrylamide Gel - methods</topic><topic>Isocitrate lyase</topic><topic>Isocitrate Lyase - analysis</topic><topic>Native sulfate-polyacrylamide gel electrophoresis</topic><topic>Protein Denaturation</topic><topic>Sodium dodecyl</topic><topic>Sodium Dodecyl Sulfate</topic><topic>Staining and Labeling - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hou, Wen-Chi</creatorcontrib><creatorcontrib>Chen, Hsien-Jung</creatorcontrib><creatorcontrib>Lin, Yaw-Huei</creatorcontrib><creatorcontrib>Chen, Yen-Chou</creatorcontrib><creatorcontrib>Yang, Ling-Ling</creatorcontrib><creatorcontrib>Lee, Mei-Hsien</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Electrophoresis</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hou, Wen-Chi</au><au>Chen, Hsien-Jung</au><au>Lin, Yaw-Huei</au><au>Chen, Yen-Chou</au><au>Yang, Ling-Ling</au><au>Lee, Mei-Hsien</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Activity staining of isocitrate lyase after electrophoresis on either native or sodium dodecyl sulfate polyacrylamide gels</atitle><jtitle>Electrophoresis</jtitle><addtitle>ELECTROPHORESIS</addtitle><date>2001-08</date><risdate>2001</risdate><volume>22</volume><issue>13</issue><spage>2653</spage><epage>2655</epage><pages>2653-2655</pages><issn>0173-0835</issn><eissn>1522-2683</eissn><abstract>Isocitrate was cleaved into succinate and glyoxylate by isocitrate lyase (ICL) in the glyoxylate cycle. 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| subjects | Acrylic Resins Activity staining polyacrylamide gel electrophoresis Electrophoresis, Polyacrylamide Gel - methods Isocitrate lyase Isocitrate Lyase - analysis Native sulfate-polyacrylamide gel electrophoresis Protein Denaturation Sodium dodecyl Sodium Dodecyl Sulfate Staining and Labeling - methods |
| title | Activity staining of isocitrate lyase after electrophoresis on either native or sodium dodecyl sulfate polyacrylamide gels |
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