A straightforward ninhydrin-based method for collagenase activity and inhibitor screening of collagenase using spectrophotometry
Currently protease assay kits, requiring substrate that is either radiolabeled or fluorescence labeled and specialized instruments, are all expensive. A simple, reliable assay of protease activity and its inhibitor screening for general laboratory is rare. Here we demonstrated a straightforward ninh...
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| Veröffentlicht in: | Analytical biochemistry 2013-06, Vol.437 (1), p.46-48 |
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| container_title | Analytical biochemistry |
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| creator | Zhang, Yanfang Fu, Yun Zhou, Sufeng Kang, Lixia Li, Changzheng |
| description | Currently protease assay kits, requiring substrate that is either radiolabeled or fluorescence labeled and specialized instruments, are all expensive. A simple, reliable assay of protease activity and its inhibitor screening for general laboratory is rare. Here we demonstrated a straightforward ninhydrin-based method for assay of collagenase activity and its inhibitor screening using spectrophotometry. In the method, without multistep sample treatments and substrate labeling, the hydrolytic products were directly traced by ninhydrin. The method is expected to be suitable for not only the assay of collagenase activity but also the others matrix metalloproteinases activities, and can be used for kinetic study. |
| doi_str_mv | 10.1016/j.ab.2013.02.030 |
| format | Article |
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A simple, reliable assay of protease activity and its inhibitor screening for general laboratory is rare. Here we demonstrated a straightforward ninhydrin-based method for assay of collagenase activity and its inhibitor screening using spectrophotometry. In the method, without multistep sample treatments and substrate labeling, the hydrolytic products were directly traced by ninhydrin. The method is expected to be suitable for not only the assay of collagenase activity but also the others matrix metalloproteinases activities, and can be used for kinetic study.</description><identifier>ISSN: 0003-2697</identifier><identifier>EISSN: 1096-0309</identifier><identifier>DOI: 10.1016/j.ab.2013.02.030</identifier><identifier>PMID: 23499966</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>collagenase ; Collagenase assay ; Collagenases - metabolism ; Drug Evaluation, Preclinical ; enzyme activity ; Enzyme Assays - methods ; fluorescence ; Hydrolysis ; Inhibitor ; Matrix Metalloproteinase Inhibitors - pharmacology ; Metallomatrix proteinase ; Ninhydrin ; Ninhydrin - chemistry ; Phenanthrolines - pharmacology ; Polyethylene Glycols - chemistry ; proteases ; radiolabeling ; screening ; Spectrophotometry ; Spectroscopy</subject><ispartof>Analytical biochemistry, 2013-06, Vol.437 (1), p.46-48</ispartof><rights>2013 Elsevier Inc.</rights><rights>Copyright © 2013 Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c440t-b751c3125bb76ed6cb096e6083e00f44e0077ca1ef890eb9225be116086931713</citedby><cites>FETCH-LOGICAL-c440t-b751c3125bb76ed6cb096e6083e00f44e0077ca1ef890eb9225be116086931713</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.ab.2013.02.030$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3548,27922,27923,45993</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23499966$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhang, Yanfang</creatorcontrib><creatorcontrib>Fu, Yun</creatorcontrib><creatorcontrib>Zhou, Sufeng</creatorcontrib><creatorcontrib>Kang, Lixia</creatorcontrib><creatorcontrib>Li, Changzheng</creatorcontrib><title>A straightforward ninhydrin-based method for collagenase activity and inhibitor screening of collagenase using spectrophotometry</title><title>Analytical biochemistry</title><addtitle>Anal Biochem</addtitle><description>Currently protease assay kits, requiring substrate that is either radiolabeled or fluorescence labeled and specialized instruments, are all expensive. A simple, reliable assay of protease activity and its inhibitor screening for general laboratory is rare. Here we demonstrated a straightforward ninhydrin-based method for assay of collagenase activity and its inhibitor screening using spectrophotometry. In the method, without multistep sample treatments and substrate labeling, the hydrolytic products were directly traced by ninhydrin. The method is expected to be suitable for not only the assay of collagenase activity but also the others matrix metalloproteinases activities, and can be used for kinetic study.</description><subject>collagenase</subject><subject>Collagenase assay</subject><subject>Collagenases - metabolism</subject><subject>Drug Evaluation, Preclinical</subject><subject>enzyme activity</subject><subject>Enzyme Assays - methods</subject><subject>fluorescence</subject><subject>Hydrolysis</subject><subject>Inhibitor</subject><subject>Matrix Metalloproteinase Inhibitors - pharmacology</subject><subject>Metallomatrix proteinase</subject><subject>Ninhydrin</subject><subject>Ninhydrin - chemistry</subject><subject>Phenanthrolines - pharmacology</subject><subject>Polyethylene Glycols - chemistry</subject><subject>proteases</subject><subject>radiolabeling</subject><subject>screening</subject><subject>Spectrophotometry</subject><subject>Spectroscopy</subject><issn>0003-2697</issn><issn>1096-0309</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kU1vGyEQhlHVqnHT3ntq99jLbgfYZUNvUdQvKVIOSc6Ij1kby15cwIl860_PWE4r9dALoOGZF3hg7D2HjgNXn9eddZ0ALjsQHUh4wRYctGppqV-yBQDIVig9nrE3pawBOO8H9ZqdCdlrrZVasN-XTanZxuWqTik_2hyaOc6rQ8hxbp0tGJot1lUKDW03Pm02dokz1Rvra3yI9dDYOTTUEl2shBSfESli2aTpH35fjsWyQ19z2q1STRScD2_Zq8luCr57ns_Z_bevd1c_2uub7z-vLq9b3_dQWzcO3EsuBudGhUF5R-9EBRcSAaa-p3EcveU4XWhApwWRyDkBSks-cnnOPp1ydzn92mOpZhuLR7rfjGlfDJdiHEU_9IpQOKE-p1IyTmaX49bmg-Fgjt7N2lhnjt4NCEOyqeXDc_rebTH8bfgjmoCPJ2CyydhljsXc31KCoj8CIflAxJcTgWThIWI2xUecPYaYyZkJKf7__Ce8mJ3F</recordid><startdate>20130601</startdate><enddate>20130601</enddate><creator>Zhang, Yanfang</creator><creator>Fu, Yun</creator><creator>Zhou, Sufeng</creator><creator>Kang, Lixia</creator><creator>Li, Changzheng</creator><general>Elsevier Inc</general><scope>FBQ</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>20130601</creationdate><title>A straightforward ninhydrin-based method for collagenase activity and inhibitor screening of collagenase using spectrophotometry</title><author>Zhang, Yanfang ; Fu, Yun ; Zhou, Sufeng ; Kang, Lixia ; Li, Changzheng</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c440t-b751c3125bb76ed6cb096e6083e00f44e0077ca1ef890eb9225be116086931713</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>collagenase</topic><topic>Collagenase assay</topic><topic>Collagenases - metabolism</topic><topic>Drug Evaluation, Preclinical</topic><topic>enzyme activity</topic><topic>Enzyme Assays - methods</topic><topic>fluorescence</topic><topic>Hydrolysis</topic><topic>Inhibitor</topic><topic>Matrix Metalloproteinase Inhibitors - pharmacology</topic><topic>Metallomatrix proteinase</topic><topic>Ninhydrin</topic><topic>Ninhydrin - chemistry</topic><topic>Phenanthrolines - pharmacology</topic><topic>Polyethylene Glycols - chemistry</topic><topic>proteases</topic><topic>radiolabeling</topic><topic>screening</topic><topic>Spectrophotometry</topic><topic>Spectroscopy</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhang, Yanfang</creatorcontrib><creatorcontrib>Fu, Yun</creatorcontrib><creatorcontrib>Zhou, Sufeng</creatorcontrib><creatorcontrib>Kang, Lixia</creatorcontrib><creatorcontrib>Li, Changzheng</creatorcontrib><collection>AGRIS</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>Analytical biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhang, Yanfang</au><au>Fu, Yun</au><au>Zhou, Sufeng</au><au>Kang, Lixia</au><au>Li, Changzheng</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A straightforward ninhydrin-based method for collagenase activity and inhibitor screening of collagenase using spectrophotometry</atitle><jtitle>Analytical biochemistry</jtitle><addtitle>Anal Biochem</addtitle><date>2013-06-01</date><risdate>2013</risdate><volume>437</volume><issue>1</issue><spage>46</spage><epage>48</epage><pages>46-48</pages><issn>0003-2697</issn><eissn>1096-0309</eissn><abstract>Currently protease assay kits, requiring substrate that is either radiolabeled or fluorescence labeled and specialized instruments, are all expensive. A simple, reliable assay of protease activity and its inhibitor screening for general laboratory is rare. Here we demonstrated a straightforward ninhydrin-based method for assay of collagenase activity and its inhibitor screening using spectrophotometry. In the method, without multistep sample treatments and substrate labeling, the hydrolytic products were directly traced by ninhydrin. The method is expected to be suitable for not only the assay of collagenase activity but also the others matrix metalloproteinases activities, and can be used for kinetic study.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>23499966</pmid><doi>10.1016/j.ab.2013.02.030</doi><tpages>3</tpages></addata></record> |
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| ispartof | Analytical biochemistry, 2013-06, Vol.437 (1), p.46-48 |
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| source | MEDLINE; ScienceDirect Journals (5 years ago - present) |
| subjects | collagenase Collagenase assay Collagenases - metabolism Drug Evaluation, Preclinical enzyme activity Enzyme Assays - methods fluorescence Hydrolysis Inhibitor Matrix Metalloproteinase Inhibitors - pharmacology Metallomatrix proteinase Ninhydrin Ninhydrin - chemistry Phenanthrolines - pharmacology Polyethylene Glycols - chemistry proteases radiolabeling screening Spectrophotometry Spectroscopy |
| title | A straightforward ninhydrin-based method for collagenase activity and inhibitor screening of collagenase using spectrophotometry |
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