An improved photochemical method for the rapid spectrophotometric detection of superoxide dismutase
A sensitive and convenient method is described for estimating superoxide dismutase activity using a photochemical augmentation procedure. This method is applicable to both liquid assays and polyacrylamide gel electropherograms. The flux of superoxide is generated by illuminating a reaction mixture c...
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| Veröffentlicht in: | Redox report : communications in free radical research 2001-01, Vol.6 (2), p.123-127 |
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| container_title | Redox report : communications in free radical research |
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| creator | Madon, P.S. |
| description | A sensitive and convenient method is described for estimating superoxide dismutase activity using a photochemical augmentation procedure. This method is applicable to both liquid assays and polyacrylamide gel electropherograms. The flux of superoxide is generated by illuminating a reaction mixture containing dianisidine and riboflavin by either a laser source or light from a fluorescent lamp. The oxidation of dianisidine, as sensitized by riboflavin, is enhanced by superoxide dismutase. The increase is linearly dependent on superoxide dismutase concentration. The photochemical reaction is allowed to proceed uninterrupted for a standardized optimum time and intensity of illumination and then terminated by addition of a buffer, 'finibuf', which stabilizes the chromophoric complex formed. This permits the spectrophotometric absorbance measurements of a number of samples collectively and also eliminates the interruption of illumination with the concomitant requirement of a spectrophotometer for constant recording of the absorbance. This method is of utility to both biochemists and clinicians. |
| doi_str_mv | 10.1179/135100001101536139 |
| format | Article |
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This method is applicable to both liquid assays and polyacrylamide gel electropherograms. The flux of superoxide is generated by illuminating a reaction mixture containing dianisidine and riboflavin by either a laser source or light from a fluorescent lamp. The oxidation of dianisidine, as sensitized by riboflavin, is enhanced by superoxide dismutase. The increase is linearly dependent on superoxide dismutase concentration. The photochemical reaction is allowed to proceed uninterrupted for a standardized optimum time and intensity of illumination and then terminated by addition of a buffer, 'finibuf', which stabilizes the chromophoric complex formed. This permits the spectrophotometric absorbance measurements of a number of samples collectively and also eliminates the interruption of illumination with the concomitant requirement of a spectrophotometer for constant recording of the absorbance. This method is of utility to both biochemists and clinicians.</description><identifier>ISSN: 1351-0002</identifier><identifier>EISSN: 1743-2928</identifier><identifier>DOI: 10.1179/135100001101536139</identifier><identifier>PMID: 11450983</identifier><language>eng</language><publisher>England: Taylor & Francis</publisher><subject>Buffers ; Chromogenic Compounds - analysis ; Dianisidine - chemistry ; Dianisidine - radiation effects ; Electrophoresis, Polyacrylamide Gel ; Fluorescence ; Lasers ; Oxidation-Reduction ; Photochemistry ; Riboflavin - chemistry ; Riboflavin - radiation effects ; Sensitivity and Specificity ; Spectrophotometry - methods ; Superoxide Dismutase - analysis ; Superoxides - metabolism</subject><ispartof>Redox report : communications in free radical research, 2001-01, Vol.6 (2), p.123-127</ispartof><rights>2001 Maney Publishing 2001</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c353t-198b6322596f31a8ca875d56d02d78c5c5f737caa92bfca0dc472be0a7b0c2b83</citedby><cites>FETCH-LOGICAL-c353t-198b6322596f31a8ca875d56d02d78c5c5f737caa92bfca0dc472be0a7b0c2b83</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11450983$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Madon, P.S.</creatorcontrib><title>An improved photochemical method for the rapid spectrophotometric detection of superoxide dismutase</title><title>Redox report : communications in free radical research</title><addtitle>Redox Rep</addtitle><description>A sensitive and convenient method is described for estimating superoxide dismutase activity using a photochemical augmentation procedure. This method is applicable to both liquid assays and polyacrylamide gel electropherograms. The flux of superoxide is generated by illuminating a reaction mixture containing dianisidine and riboflavin by either a laser source or light from a fluorescent lamp. The oxidation of dianisidine, as sensitized by riboflavin, is enhanced by superoxide dismutase. The increase is linearly dependent on superoxide dismutase concentration. The photochemical reaction is allowed to proceed uninterrupted for a standardized optimum time and intensity of illumination and then terminated by addition of a buffer, 'finibuf', which stabilizes the chromophoric complex formed. This permits the spectrophotometric absorbance measurements of a number of samples collectively and also eliminates the interruption of illumination with the concomitant requirement of a spectrophotometer for constant recording of the absorbance. This method is of utility to both biochemists and clinicians.</description><subject>Buffers</subject><subject>Chromogenic Compounds - analysis</subject><subject>Dianisidine - chemistry</subject><subject>Dianisidine - radiation effects</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Fluorescence</subject><subject>Lasers</subject><subject>Oxidation-Reduction</subject><subject>Photochemistry</subject><subject>Riboflavin - chemistry</subject><subject>Riboflavin - radiation effects</subject><subject>Sensitivity and Specificity</subject><subject>Spectrophotometry - methods</subject><subject>Superoxide Dismutase - analysis</subject><subject>Superoxides - metabolism</subject><issn>1351-0002</issn><issn>1743-2928</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kEtLxDAQgIMo7rr6BzxITt6qeTRNevCwLL5gwYueS5oHG2mbmqTq_nuju-BBcC4zDN_MMB8A5xhdYczra0wZRjkwRpjRCtP6AMwxL2lBaiIOc52BIgNkBk5ifM0VrWpxDGYYlwzVgs6BWg7Q9WPw70bDceOTVxvTOyU72Ju08RpaH2DaGBjk6DSMo1Ep-B8yA8EpqE3KPecH6C2M02iC_3TaQO1iPyUZzSk4srKL5myfF-Dl7vZ59VCsn-4fV8t1oSijqcC1aCtKCKsrS7EUSgrONKs0IpoLxRSznHIlZU1aqyTSquSkNUjyFinSCroAl7u9-Z23ycTU9C4q03VyMH6KDc-eygpXGSQ7UAUfYzC2GYPrZdg2GDXfapu_avPQxX771PZG_47sXWbgZge4ITvr5YcPnW6S3HY-2CAH5WJD_znwBQ8iiOA</recordid><startdate>20010101</startdate><enddate>20010101</enddate><creator>Madon, P.S.</creator><general>Taylor & Francis</general><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>20010101</creationdate><title>An improved photochemical method for the rapid spectrophotometric detection of superoxide dismutase</title><author>Madon, P.S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c353t-198b6322596f31a8ca875d56d02d78c5c5f737caa92bfca0dc472be0a7b0c2b83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>Buffers</topic><topic>Chromogenic Compounds - analysis</topic><topic>Dianisidine - chemistry</topic><topic>Dianisidine - radiation effects</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Fluorescence</topic><topic>Lasers</topic><topic>Oxidation-Reduction</topic><topic>Photochemistry</topic><topic>Riboflavin - chemistry</topic><topic>Riboflavin - radiation effects</topic><topic>Sensitivity and Specificity</topic><topic>Spectrophotometry - methods</topic><topic>Superoxide Dismutase - analysis</topic><topic>Superoxides - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Madon, P.S.</creatorcontrib><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>Redox report : communications in free radical research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Madon, P.S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>An improved photochemical method for the rapid spectrophotometric detection of superoxide dismutase</atitle><jtitle>Redox report : communications in free radical research</jtitle><addtitle>Redox Rep</addtitle><date>2001-01-01</date><risdate>2001</risdate><volume>6</volume><issue>2</issue><spage>123</spage><epage>127</epage><pages>123-127</pages><issn>1351-0002</issn><eissn>1743-2928</eissn><abstract>A sensitive and convenient method is described for estimating superoxide dismutase activity using a photochemical augmentation procedure. This method is applicable to both liquid assays and polyacrylamide gel electropherograms. The flux of superoxide is generated by illuminating a reaction mixture containing dianisidine and riboflavin by either a laser source or light from a fluorescent lamp. The oxidation of dianisidine, as sensitized by riboflavin, is enhanced by superoxide dismutase. The increase is linearly dependent on superoxide dismutase concentration. The photochemical reaction is allowed to proceed uninterrupted for a standardized optimum time and intensity of illumination and then terminated by addition of a buffer, 'finibuf', which stabilizes the chromophoric complex formed. This permits the spectrophotometric absorbance measurements of a number of samples collectively and also eliminates the interruption of illumination with the concomitant requirement of a spectrophotometer for constant recording of the absorbance. 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| fulltext | fulltext |
| identifier | ISSN: 1351-0002 |
| ispartof | Redox report : communications in free radical research, 2001-01, Vol.6 (2), p.123-127 |
| issn | 1351-0002 1743-2928 |
| language | eng |
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| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
| subjects | Buffers Chromogenic Compounds - analysis Dianisidine - chemistry Dianisidine - radiation effects Electrophoresis, Polyacrylamide Gel Fluorescence Lasers Oxidation-Reduction Photochemistry Riboflavin - chemistry Riboflavin - radiation effects Sensitivity and Specificity Spectrophotometry - methods Superoxide Dismutase - analysis Superoxides - metabolism |
| title | An improved photochemical method for the rapid spectrophotometric detection of superoxide dismutase |
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