Generation of a monoclonal antibody against the myelin protein CNP (2',3'-cyclic nucleotide 3'-phosphodiesterase) suitable for biochemical and for immunohistochemical investigations of CNP
The functional role of CNP (2',3'-cyclic nucleotide 3'-phosphodiesterase), a minor component of central and peripheral myelin is still unclear. Here we describe preparation of a monoclonal antibody directed against CNP. The antibody, of the immunoglobulin IgG1 type, raised with a basi...
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Veröffentlicht in: | Neurochemical research 1994-12, Vol.19 (12), p.1479-1485 |
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description | The functional role of CNP (2',3'-cyclic nucleotide 3'-phosphodiesterase), a minor component of central and peripheral myelin is still unclear. Here we describe preparation of a monoclonal antibody directed against CNP. The antibody, of the immunoglobulin IgG1 type, raised with a basic 46 kDa membrane-associated protein solubilized from pig cerebellar membranes, can be used to detect immunoreactivity in solubilized brain homogenates from pig, mouse, rat, sheep, cow and man, in cerebrum and cerebellum, but not in other tissues such as liver, skeletal and heart muscle. The antibody recognizes the CNP doublet band and shows no cross-reactivity with any of the other brain proteins solubilized. In tissue sections from paraformaldehyde-fixed rat brain the antigen was localized in oligodendrocytes. In cultured glial cells from newborn mice the antibody stained cells which were identified as oligodendrocytes by co-localization of myelin basic protein. Even cells from a C6 rat glioma cell line, which contain very little of CNP, were labeled by the monoclonal antibody. Thus the monoclonal antibody recognizing CNP from several species is suitable for immunocytochemical investigations and also for biochemical studies of CNP, since the antibody has been employed for immunoprecipitation and immunopurification of CNP in crude brain homogenates. |
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Here we describe preparation of a monoclonal antibody directed against CNP. The antibody, of the immunoglobulin IgG1 type, raised with a basic 46 kDa membrane-associated protein solubilized from pig cerebellar membranes, can be used to detect immunoreactivity in solubilized brain homogenates from pig, mouse, rat, sheep, cow and man, in cerebrum and cerebellum, but not in other tissues such as liver, skeletal and heart muscle. The antibody recognizes the CNP doublet band and shows no cross-reactivity with any of the other brain proteins solubilized. In tissue sections from paraformaldehyde-fixed rat brain the antigen was localized in oligodendrocytes. In cultured glial cells from newborn mice the antibody stained cells which were identified as oligodendrocytes by co-localization of myelin basic protein. Even cells from a C6 rat glioma cell line, which contain very little of CNP, were labeled by the monoclonal antibody. Thus the monoclonal antibody recognizing CNP from several species is suitable for immunocytochemical investigations and also for biochemical studies of CNP, since the antibody has been employed for immunoprecipitation and immunopurification of CNP in crude brain homogenates.</description><identifier>ISSN: 0364-3190</identifier><identifier>EISSN: 1573-6903</identifier><identifier>DOI: 10.1007/BF00968994</identifier><identifier>PMID: 7877717</identifier><identifier>CODEN: NEREDZ</identifier><language>eng</language><publisher>New York, NY: Springer</publisher><subject>2',3'-Cyclic-Nucleotide Phosphodiesterases - analysis ; 2',3'-Cyclic-Nucleotide Phosphodiesterases - immunology ; Animals ; Antibodies, Monoclonal - biosynthesis ; Antibodies, Monoclonal - immunology ; Antibody Specificity ; Biological and medical sciences ; Blotting, Western ; Brain - enzymology ; Cattle ; Cell Membrane - enzymology ; Cells, Cultured ; Cerebellum - enzymology ; Fluorescent Antibody Technique ; Fundamental and applied biological sciences. Psychology ; Humans ; Immunoglobulin G - immunology ; Immunohistochemistry ; Isolated neuron and nerve. Neuroglia ; Mice ; Mice, Inbred BALB C ; Neuroglia - enzymology ; Rats ; Rodentia ; Sheep ; Swine ; Tissue Distribution ; Vertebrates: nervous system and sense organs</subject><ispartof>Neurochemical research, 1994-12, Vol.19 (12), p.1479-1485</ispartof><rights>1995 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c342t-85d38c9d769b102001f9749984d01a86850b46c8928d47414f17eb8db6e6fbde3</citedby><cites>FETCH-LOGICAL-c342t-85d38c9d769b102001f9749984d01a86850b46c8928d47414f17eb8db6e6fbde3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,782,786,27931,27932</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=3391628$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7877717$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>REISER, G</creatorcontrib><creatorcontrib>KUNZELMANN, U</creatorcontrib><creatorcontrib>STEINHILBER, G</creatorcontrib><creatorcontrib>BINMÖLLER, F.-J</creatorcontrib><title>Generation of a monoclonal antibody against the myelin protein CNP (2',3'-cyclic nucleotide 3'-phosphodiesterase) suitable for biochemical and for immunohistochemical investigations of CNP</title><title>Neurochemical research</title><addtitle>Neurochem Res</addtitle><description>The functional role of CNP (2',3'-cyclic nucleotide 3'-phosphodiesterase), a minor component of central and peripheral myelin is still unclear. Here we describe preparation of a monoclonal antibody directed against CNP. The antibody, of the immunoglobulin IgG1 type, raised with a basic 46 kDa membrane-associated protein solubilized from pig cerebellar membranes, can be used to detect immunoreactivity in solubilized brain homogenates from pig, mouse, rat, sheep, cow and man, in cerebrum and cerebellum, but not in other tissues such as liver, skeletal and heart muscle. The antibody recognizes the CNP doublet band and shows no cross-reactivity with any of the other brain proteins solubilized. In tissue sections from paraformaldehyde-fixed rat brain the antigen was localized in oligodendrocytes. In cultured glial cells from newborn mice the antibody stained cells which were identified as oligodendrocytes by co-localization of myelin basic protein. Even cells from a C6 rat glioma cell line, which contain very little of CNP, were labeled by the monoclonal antibody. Thus the monoclonal antibody recognizing CNP from several species is suitable for immunocytochemical investigations and also for biochemical studies of CNP, since the antibody has been employed for immunoprecipitation and immunopurification of CNP in crude brain homogenates.</description><subject>2',3'-Cyclic-Nucleotide Phosphodiesterases - analysis</subject><subject>2',3'-Cyclic-Nucleotide Phosphodiesterases - immunology</subject><subject>Animals</subject><subject>Antibodies, Monoclonal - biosynthesis</subject><subject>Antibodies, Monoclonal - immunology</subject><subject>Antibody Specificity</subject><subject>Biological and medical sciences</subject><subject>Blotting, Western</subject><subject>Brain - enzymology</subject><subject>Cattle</subject><subject>Cell Membrane - enzymology</subject><subject>Cells, Cultured</subject><subject>Cerebellum - enzymology</subject><subject>Fluorescent Antibody Technique</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Humans</subject><subject>Immunoglobulin G - immunology</subject><subject>Immunohistochemistry</subject><subject>Isolated neuron and nerve. Neuroglia</subject><subject>Mice</subject><subject>Mice, Inbred BALB C</subject><subject>Neuroglia - enzymology</subject><subject>Rats</subject><subject>Rodentia</subject><subject>Sheep</subject><subject>Swine</subject><subject>Tissue Distribution</subject><subject>Vertebrates: nervous system and sense organs</subject><issn>0364-3190</issn><issn>1573-6903</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU9v1DAQxS0EKkvLhTuSD4hCRcCOHf85woq2SFXhAOfIsSddI8fexg7Sfjc-HO521R45WCPN-2lm_B5Cryj5SAmRn76cE6KF0po_QSvaSdYITdhTtCJM8IZRTZ6jFzn_JqTiLT1CR1JJKalcob8XEGE2xaeI04gNnlJMNqRoAjax-CG5HTY3xsdccNkAnnYQfMTbORWodX39A79rTz-w08bubPAWx8UGSMU7wLW53aRcn_OQS92T4T3Oiy9mCIDHNOPBJ7uBydv9Prfv-WlaYtr4XB41H__UCf5mf2m-O7VuPkHPRhMyvDzUY_Tr_OvP9WVz9f3i2_rzVWMZb0ujOseU1U4KPVDSVhdGLbnWijtCjRKqIwMXVulWOS455SOVMCg3CBDj4IAdo7f3c-uvb5d6Rz_5bCEEEyEtua9eKtV17X9BKkTbccIreHYP2jnlPMPYb2c_mXnXU9LfZdo_Zlrh14epyzCBe0APIVb9zUE3uXo1ziZanx8wxjQVrWL_AMbGqpQ</recordid><startdate>19941201</startdate><enddate>19941201</enddate><creator>REISER, G</creator><creator>KUNZELMANN, U</creator><creator>STEINHILBER, G</creator><creator>BINMÖLLER, F.-J</creator><general>Springer</general><scope>IQODW</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>7TK</scope><scope>7X8</scope></search><sort><creationdate>19941201</creationdate><title>Generation of a monoclonal antibody against the myelin protein CNP (2',3'-cyclic nucleotide 3'-phosphodiesterase) suitable for biochemical and for immunohistochemical investigations of CNP</title><author>REISER, G ; KUNZELMANN, U ; STEINHILBER, G ; BINMÖLLER, F.-J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c342t-85d38c9d769b102001f9749984d01a86850b46c8928d47414f17eb8db6e6fbde3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>2',3'-Cyclic-Nucleotide Phosphodiesterases - analysis</topic><topic>2',3'-Cyclic-Nucleotide Phosphodiesterases - immunology</topic><topic>Animals</topic><topic>Antibodies, Monoclonal - biosynthesis</topic><topic>Antibodies, Monoclonal - immunology</topic><topic>Antibody Specificity</topic><topic>Biological and medical sciences</topic><topic>Blotting, Western</topic><topic>Brain - enzymology</topic><topic>Cattle</topic><topic>Cell Membrane - enzymology</topic><topic>Cells, Cultured</topic><topic>Cerebellum - enzymology</topic><topic>Fluorescent Antibody Technique</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Humans</topic><topic>Immunoglobulin G - immunology</topic><topic>Immunohistochemistry</topic><topic>Isolated neuron and nerve. Neuroglia</topic><topic>Mice</topic><topic>Mice, Inbred BALB C</topic><topic>Neuroglia - enzymology</topic><topic>Rats</topic><topic>Rodentia</topic><topic>Sheep</topic><topic>Swine</topic><topic>Tissue Distribution</topic><topic>Vertebrates: nervous system and sense organs</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>REISER, G</creatorcontrib><creatorcontrib>KUNZELMANN, U</creatorcontrib><creatorcontrib>STEINHILBER, G</creatorcontrib><creatorcontrib>BINMÖLLER, F.-J</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Neurosciences Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Neurochemical research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>REISER, G</au><au>KUNZELMANN, U</au><au>STEINHILBER, G</au><au>BINMÖLLER, F.-J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Generation of a monoclonal antibody against the myelin protein CNP (2',3'-cyclic nucleotide 3'-phosphodiesterase) suitable for biochemical and for immunohistochemical investigations of CNP</atitle><jtitle>Neurochemical research</jtitle><addtitle>Neurochem Res</addtitle><date>1994-12-01</date><risdate>1994</risdate><volume>19</volume><issue>12</issue><spage>1479</spage><epage>1485</epage><pages>1479-1485</pages><issn>0364-3190</issn><eissn>1573-6903</eissn><coden>NEREDZ</coden><abstract>The functional role of CNP (2',3'-cyclic nucleotide 3'-phosphodiesterase), a minor component of central and peripheral myelin is still unclear. Here we describe preparation of a monoclonal antibody directed against CNP. The antibody, of the immunoglobulin IgG1 type, raised with a basic 46 kDa membrane-associated protein solubilized from pig cerebellar membranes, can be used to detect immunoreactivity in solubilized brain homogenates from pig, mouse, rat, sheep, cow and man, in cerebrum and cerebellum, but not in other tissues such as liver, skeletal and heart muscle. The antibody recognizes the CNP doublet band and shows no cross-reactivity with any of the other brain proteins solubilized. In tissue sections from paraformaldehyde-fixed rat brain the antigen was localized in oligodendrocytes. In cultured glial cells from newborn mice the antibody stained cells which were identified as oligodendrocytes by co-localization of myelin basic protein. Even cells from a C6 rat glioma cell line, which contain very little of CNP, were labeled by the monoclonal antibody. Thus the monoclonal antibody recognizing CNP from several species is suitable for immunocytochemical investigations and also for biochemical studies of CNP, since the antibody has been employed for immunoprecipitation and immunopurification of CNP in crude brain homogenates.</abstract><cop>New York, NY</cop><pub>Springer</pub><pmid>7877717</pmid><doi>10.1007/BF00968994</doi><tpages>7</tpages></addata></record> |
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subjects | 2',3'-Cyclic-Nucleotide Phosphodiesterases - analysis 2',3'-Cyclic-Nucleotide Phosphodiesterases - immunology Animals Antibodies, Monoclonal - biosynthesis Antibodies, Monoclonal - immunology Antibody Specificity Biological and medical sciences Blotting, Western Brain - enzymology Cattle Cell Membrane - enzymology Cells, Cultured Cerebellum - enzymology Fluorescent Antibody Technique Fundamental and applied biological sciences. Psychology Humans Immunoglobulin G - immunology Immunohistochemistry Isolated neuron and nerve. Neuroglia Mice Mice, Inbred BALB C Neuroglia - enzymology Rats Rodentia Sheep Swine Tissue Distribution Vertebrates: nervous system and sense organs |
title | Generation of a monoclonal antibody against the myelin protein CNP (2',3'-cyclic nucleotide 3'-phosphodiesterase) suitable for biochemical and for immunohistochemical investigations of CNP |
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