Abstract
We immunized rats with recombinant murine osteopontin protein and obtained four monoclonal antibodies recognizing distinct epitopes of murine osteopontin. OPN1.2 recognized the amino-terminal half of OPN, while OPN2.2, OPN2.3, and OPN3.1 recognized the carboxy-terminal half of OPN. The epitope recognized by OPN2.2 was destroyed by further cleavage of the carboxy half of OPN. The epitope recognized by OPN2.3 was located in the amino-terminal end of the carboxy half of OPN, whereas that recognized by OPN3.1 was located in the carboxy-terminal end of the carboxy half of OPN. OPN1.2 and OPN2.2 recognized thrombin-cleaved osteopontin, whereas thrombin-cleaved osteopontin was not recognized by OPN2.3 and OPN3.1. Thus, these monoclonal antibodies will be useful in structure/function studies of the role of osteopontin in murine models of disease.
Copyright 1999 Academic Press.
MeSH terms
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Animals
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Antibodies, Monoclonal / immunology*
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Antibody Specificity
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Blotting, Western
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Calcification, Physiologic
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Epitopes / analysis
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Epitopes / immunology*
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Immunohistochemistry
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Mice
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Mice, Inbred BALB C
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Molecular Weight
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Mutation
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Myocardium / chemistry
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Myocardium / cytology
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Oligopeptides / genetics
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Oligopeptides / immunology
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Osteopontin
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Peptide Fragments / genetics
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Peptide Fragments / immunology
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Protein Conformation
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Rats
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / immunology
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Recombinant Fusion Proteins / metabolism
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Sequence Deletion
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Sialoglycoproteins / analysis
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Sialoglycoproteins / chemistry
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Sialoglycoproteins / genetics
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Sialoglycoproteins / immunology*
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Thrombin / metabolism
Substances
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Antibodies, Monoclonal
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Epitopes
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Oligopeptides
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Peptide Fragments
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Recombinant Fusion Proteins
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Sialoglycoproteins
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Spp1 protein, mouse
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Spp1 protein, rat
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Osteopontin
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arginyl-glycyl-aspartic acid
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Thrombin