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
Hauptverfasser: REISER, G, KUNZELMANN, U, STEINHILBER, G, BINMÖLLER, F.-J
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container_issue 12
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container_title Neurochemical research
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creator REISER, G
KUNZELMANN, U
STEINHILBER, G
BINMÖLLER, F.-J
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. 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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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