The Experts below are selected from a list of 48 Experts worldwide ranked by ideXlab platform
Peter J. Houghton - One of the best experts on this subject based on the ideXlab platform.
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Monoclonal antibodies to the myogenic regulatory Protein MyoD1: epitope mapping and diagnostic utility.
Cancer research, 1992Co-Authors: P Dias, David M. Parham, David N. Shapiro, Stephen J. Tapscott, Peter J. HoughtonAbstract:Monoclonal antibodies (MoAbs) were developed against recombinant wild-type murine MyoD1 Protein. Each of 4 MoAbs was immunologically reactive with recombinant MyoD1 Protein by enzyme-linked immunosorbent assay, and each specifically stained the nuclei of myogenic cells. Epitopes were mapped using fusion Protein constructs with specific deletions of defined regions of the MyoD1 molecule. MoAb 5.2F recognized an epitope in the amino terminal region between amino acid residues (AAR) 3 and 56, whereas epitopes for MoAbs 1.1A, 5.4G, and 5.8A were in the carboxyl terminus (AAR 167-318) of the MyoD1 Protein. The epitope for MoAb 5.8A was further delineated to AAR 170-209 by Western analysis and immunoprecipitation of in vivo transcribed and translated MyoD1 Protein having specific deletions in the carboxyl terminus. The 5.8A epitope was ultimately localized to the region between AAR 180 and 189 of the Protein by enzyme-linked immunosorbent assay using 10-amino acid residue synthetic peptides. This sequence is apparently unique to MyoD1 and has little homology to other myogenic regulatory Proteins (myogenin, Myf5, Myf6, and MRF4). Transfection of cDNA for murine MyoD1 into a nonmuscle cell line conferred 5.8A reactivity, confirming the specificity of this reagent. MoAb 5.8A was then used to examine the expression of MyoD1 in normal and malignant human tissues. MyoD1 was not detected in any normal adult tissue but was detected in 25 of 25 histologically confirmed rhabdomyosarcomas. Staining was localized to the nucleus and showed marked heterogeneity between cells as well as differential staining within nuclei. Specific subcellular localization of 5.8A was further determined by immunoelectron microscopy, where antibody was found to localize to electron-dense areas, more frequently associated with the nuclear submembranous region. In addition to rhabdomyosarcomas, MoAb 5.8A stained 2 of 5 Wilms' tumors and one ectomesenchymoma, neoplasms known to contain myogenic elements. The 5.8A reagent was also of value in the accurate histopathological classification of 2 of 4 tumors previously diagnosed as extraosseous Ewing's sarcoma and 2 of 3 tumors diagnosed as undifferentiated sarcomas.
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monoclonal antibodies to the myogenic regulatory Protein MyoD1 epitope mapping and diagnostic utility
Cancer Research, 1992Co-Authors: P Dias, David M. Parham, David N. Shapiro, Stephen J. Tapscott, Peter J. HoughtonAbstract:Abstract Monoclonal antibodies (MoAbs) were developed against recombinant wild-type murine MyoD1 Protein. Each of 4 MoAbs was immunologically reactive with recombinant MyoD1 Protein by enzyme-linked immunosorbent assay, and each specifically stained the nuclei of myogenic cells. Epitopes were mapped using fusion Protein constructs with specific deletions of defined regions of the MyoD1 molecule. MoAb 5.2F recognized an epitope in the amino terminal region between amino acid residues (AAR) 3 and 56, whereas epitopes for MoAbs 1.1A, 5.4G, and 5.8A were in the carboxyl terminus (AAR 167–318) of the MyoD1 Protein. The epitope for MoAb 5.8A was further delineated to AAR 170–209 by Western analysis and immunoprecipitation of in vitro transcribed and translated MyoDl Protein having specific deletions in the carboxyl terminus. The 5.8A epitope was ultimately localized to the region between AAR 180 and 189 of the Protein by enzyme-linked immunosorbent assay using 10-amino acid residue synthetic peptides. This sequence is apparently unique to MyoD1 and has little homology to other myogenic regulatory Proteins (myogenin, Myf5, Myf6, and MRF4). Transfection of cDNA for murine MyoDl into a nonmuscle cell line conferred 5.8A reactivity, confirming the specificity of this reagent. MoAb 5.8A was then used to examine the expression of MyoD1 in normal and malignant human tissues. MyoD1 was not detected in any normal adult tissue but was detected in 25 of 25 histologically confirmed rhabdomyosarcomas. Staining was localized to the nucleus and showed marked heterogeneity between cells as well as differential staining within nuclei. Specific subcellular localization of 5.8A was further determined by immunoelectron microscopy, where antibody was found to localize to electron-dense areas, more frequently associated with the nuclear submembranous region. In addition to rhabdomyosarcomas, MoAb 5.8A stained 2 of 5 Wilms9 tumors and one ectomesenchymoma, neoplasms known to contain myogenic elements. The 5.8A reagent was also of value in the accurate histopathological classification of 2 of 4 tumors previously diagnosed as extraosseous Ewing9s sarcoma and 2 of 3 tumors diaonnsed as undifferenriated sarCOmas. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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Desmin positivity in primitive neuroectodermal tumors of childhood.
The American journal of surgical pathology, 1992Co-Authors: David M. Parham, Peter Dias, David R. Kelly, Joe C. Rutledge, Peter J. HoughtonAbstract:In this report, we describe two rosette-forming primitive neuroectodermal tumors that were found to contain desmin by both immunohistochemistry and Western blotting. Electron microscopy on both cases was consistent with primitive neuroectodermal tumors and revealed that the tumor cells contained cytoplasmic bundles of intermediate filaments. In both cases, studies for MyoD1 Protein using immunohistochemistry and Western blotting were negative. Thus, the detection of desmin in a pediatric neoplasm does not absolutely exclude the diagnosis of primitive neuroectodermal tumor and should not be considered as prima facie evidence that a small-cell tumor is a rhabdomyosarcoma.
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MyoDl Protein Expression in Alveolar Soft Part Sarcoma As Confirmatory Evidence of Its Skeletal Muscle Nature
The American journal of surgical pathology, 1991Co-Authors: Juan Rosai, David M. Parham, David N. Shapiro, Peter Dias, Peter J. HoughtonAbstract:A typical case of alveolar soft part sarcoma was found to express in a strong and widespread fashion the marker MyoD1 Protein. This nuclear phosphoProtein is the product of a regulatory gene that controls the commitment of a cell to myogenic lineage; it therefore provides strong and perhaps definitive evidence in support of the skeletal muscle nature of this enigmatic neoplasm.
David M. Parham - One of the best experts on this subject based on the ideXlab platform.
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Monoclonal antibodies to the myogenic regulatory Protein MyoD1: epitope mapping and diagnostic utility.
Cancer research, 1992Co-Authors: P Dias, David M. Parham, David N. Shapiro, Stephen J. Tapscott, Peter J. HoughtonAbstract:Monoclonal antibodies (MoAbs) were developed against recombinant wild-type murine MyoD1 Protein. Each of 4 MoAbs was immunologically reactive with recombinant MyoD1 Protein by enzyme-linked immunosorbent assay, and each specifically stained the nuclei of myogenic cells. Epitopes were mapped using fusion Protein constructs with specific deletions of defined regions of the MyoD1 molecule. MoAb 5.2F recognized an epitope in the amino terminal region between amino acid residues (AAR) 3 and 56, whereas epitopes for MoAbs 1.1A, 5.4G, and 5.8A were in the carboxyl terminus (AAR 167-318) of the MyoD1 Protein. The epitope for MoAb 5.8A was further delineated to AAR 170-209 by Western analysis and immunoprecipitation of in vivo transcribed and translated MyoD1 Protein having specific deletions in the carboxyl terminus. The 5.8A epitope was ultimately localized to the region between AAR 180 and 189 of the Protein by enzyme-linked immunosorbent assay using 10-amino acid residue synthetic peptides. This sequence is apparently unique to MyoD1 and has little homology to other myogenic regulatory Proteins (myogenin, Myf5, Myf6, and MRF4). Transfection of cDNA for murine MyoD1 into a nonmuscle cell line conferred 5.8A reactivity, confirming the specificity of this reagent. MoAb 5.8A was then used to examine the expression of MyoD1 in normal and malignant human tissues. MyoD1 was not detected in any normal adult tissue but was detected in 25 of 25 histologically confirmed rhabdomyosarcomas. Staining was localized to the nucleus and showed marked heterogeneity between cells as well as differential staining within nuclei. Specific subcellular localization of 5.8A was further determined by immunoelectron microscopy, where antibody was found to localize to electron-dense areas, more frequently associated with the nuclear submembranous region. In addition to rhabdomyosarcomas, MoAb 5.8A stained 2 of 5 Wilms' tumors and one ectomesenchymoma, neoplasms known to contain myogenic elements. The 5.8A reagent was also of value in the accurate histopathological classification of 2 of 4 tumors previously diagnosed as extraosseous Ewing's sarcoma and 2 of 3 tumors diagnosed as undifferentiated sarcomas.
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monoclonal antibodies to the myogenic regulatory Protein MyoD1 epitope mapping and diagnostic utility
Cancer Research, 1992Co-Authors: P Dias, David M. Parham, David N. Shapiro, Stephen J. Tapscott, Peter J. HoughtonAbstract:Abstract Monoclonal antibodies (MoAbs) were developed against recombinant wild-type murine MyoD1 Protein. Each of 4 MoAbs was immunologically reactive with recombinant MyoD1 Protein by enzyme-linked immunosorbent assay, and each specifically stained the nuclei of myogenic cells. Epitopes were mapped using fusion Protein constructs with specific deletions of defined regions of the MyoD1 molecule. MoAb 5.2F recognized an epitope in the amino terminal region between amino acid residues (AAR) 3 and 56, whereas epitopes for MoAbs 1.1A, 5.4G, and 5.8A were in the carboxyl terminus (AAR 167–318) of the MyoD1 Protein. The epitope for MoAb 5.8A was further delineated to AAR 170–209 by Western analysis and immunoprecipitation of in vitro transcribed and translated MyoDl Protein having specific deletions in the carboxyl terminus. The 5.8A epitope was ultimately localized to the region between AAR 180 and 189 of the Protein by enzyme-linked immunosorbent assay using 10-amino acid residue synthetic peptides. This sequence is apparently unique to MyoD1 and has little homology to other myogenic regulatory Proteins (myogenin, Myf5, Myf6, and MRF4). Transfection of cDNA for murine MyoDl into a nonmuscle cell line conferred 5.8A reactivity, confirming the specificity of this reagent. MoAb 5.8A was then used to examine the expression of MyoD1 in normal and malignant human tissues. MyoD1 was not detected in any normal adult tissue but was detected in 25 of 25 histologically confirmed rhabdomyosarcomas. Staining was localized to the nucleus and showed marked heterogeneity between cells as well as differential staining within nuclei. Specific subcellular localization of 5.8A was further determined by immunoelectron microscopy, where antibody was found to localize to electron-dense areas, more frequently associated with the nuclear submembranous region. In addition to rhabdomyosarcomas, MoAb 5.8A stained 2 of 5 Wilms9 tumors and one ectomesenchymoma, neoplasms known to contain myogenic elements. The 5.8A reagent was also of value in the accurate histopathological classification of 2 of 4 tumors previously diagnosed as extraosseous Ewing9s sarcoma and 2 of 3 tumors diaonnsed as undifferenriated sarCOmas. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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Desmin positivity in primitive neuroectodermal tumors of childhood.
The American journal of surgical pathology, 1992Co-Authors: David M. Parham, Peter Dias, David R. Kelly, Joe C. Rutledge, Peter J. HoughtonAbstract:In this report, we describe two rosette-forming primitive neuroectodermal tumors that were found to contain desmin by both immunohistochemistry and Western blotting. Electron microscopy on both cases was consistent with primitive neuroectodermal tumors and revealed that the tumor cells contained cytoplasmic bundles of intermediate filaments. In both cases, studies for MyoD1 Protein using immunohistochemistry and Western blotting were negative. Thus, the detection of desmin in a pediatric neoplasm does not absolutely exclude the diagnosis of primitive neuroectodermal tumor and should not be considered as prima facie evidence that a small-cell tumor is a rhabdomyosarcoma.
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MyoDl Protein Expression in Alveolar Soft Part Sarcoma As Confirmatory Evidence of Its Skeletal Muscle Nature
The American journal of surgical pathology, 1991Co-Authors: Juan Rosai, David M. Parham, David N. Shapiro, Peter Dias, Peter J. HoughtonAbstract:A typical case of alveolar soft part sarcoma was found to express in a strong and widespread fashion the marker MyoD1 Protein. This nuclear phosphoProtein is the product of a regulatory gene that controls the commitment of a cell to myogenic lineage; it therefore provides strong and perhaps definitive evidence in support of the skeletal muscle nature of this enigmatic neoplasm.
David N. Shapiro - One of the best experts on this subject based on the ideXlab platform.
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Monoclonal antibodies to the myogenic regulatory Protein MyoD1: epitope mapping and diagnostic utility.
Cancer research, 1992Co-Authors: P Dias, David M. Parham, David N. Shapiro, Stephen J. Tapscott, Peter J. HoughtonAbstract:Monoclonal antibodies (MoAbs) were developed against recombinant wild-type murine MyoD1 Protein. Each of 4 MoAbs was immunologically reactive with recombinant MyoD1 Protein by enzyme-linked immunosorbent assay, and each specifically stained the nuclei of myogenic cells. Epitopes were mapped using fusion Protein constructs with specific deletions of defined regions of the MyoD1 molecule. MoAb 5.2F recognized an epitope in the amino terminal region between amino acid residues (AAR) 3 and 56, whereas epitopes for MoAbs 1.1A, 5.4G, and 5.8A were in the carboxyl terminus (AAR 167-318) of the MyoD1 Protein. The epitope for MoAb 5.8A was further delineated to AAR 170-209 by Western analysis and immunoprecipitation of in vivo transcribed and translated MyoD1 Protein having specific deletions in the carboxyl terminus. The 5.8A epitope was ultimately localized to the region between AAR 180 and 189 of the Protein by enzyme-linked immunosorbent assay using 10-amino acid residue synthetic peptides. This sequence is apparently unique to MyoD1 and has little homology to other myogenic regulatory Proteins (myogenin, Myf5, Myf6, and MRF4). Transfection of cDNA for murine MyoD1 into a nonmuscle cell line conferred 5.8A reactivity, confirming the specificity of this reagent. MoAb 5.8A was then used to examine the expression of MyoD1 in normal and malignant human tissues. MyoD1 was not detected in any normal adult tissue but was detected in 25 of 25 histologically confirmed rhabdomyosarcomas. Staining was localized to the nucleus and showed marked heterogeneity between cells as well as differential staining within nuclei. Specific subcellular localization of 5.8A was further determined by immunoelectron microscopy, where antibody was found to localize to electron-dense areas, more frequently associated with the nuclear submembranous region. In addition to rhabdomyosarcomas, MoAb 5.8A stained 2 of 5 Wilms' tumors and one ectomesenchymoma, neoplasms known to contain myogenic elements. The 5.8A reagent was also of value in the accurate histopathological classification of 2 of 4 tumors previously diagnosed as extraosseous Ewing's sarcoma and 2 of 3 tumors diagnosed as undifferentiated sarcomas.
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monoclonal antibodies to the myogenic regulatory Protein MyoD1 epitope mapping and diagnostic utility
Cancer Research, 1992Co-Authors: P Dias, David M. Parham, David N. Shapiro, Stephen J. Tapscott, Peter J. HoughtonAbstract:Abstract Monoclonal antibodies (MoAbs) were developed against recombinant wild-type murine MyoD1 Protein. Each of 4 MoAbs was immunologically reactive with recombinant MyoD1 Protein by enzyme-linked immunosorbent assay, and each specifically stained the nuclei of myogenic cells. Epitopes were mapped using fusion Protein constructs with specific deletions of defined regions of the MyoD1 molecule. MoAb 5.2F recognized an epitope in the amino terminal region between amino acid residues (AAR) 3 and 56, whereas epitopes for MoAbs 1.1A, 5.4G, and 5.8A were in the carboxyl terminus (AAR 167–318) of the MyoD1 Protein. The epitope for MoAb 5.8A was further delineated to AAR 170–209 by Western analysis and immunoprecipitation of in vitro transcribed and translated MyoDl Protein having specific deletions in the carboxyl terminus. The 5.8A epitope was ultimately localized to the region between AAR 180 and 189 of the Protein by enzyme-linked immunosorbent assay using 10-amino acid residue synthetic peptides. This sequence is apparently unique to MyoD1 and has little homology to other myogenic regulatory Proteins (myogenin, Myf5, Myf6, and MRF4). Transfection of cDNA for murine MyoDl into a nonmuscle cell line conferred 5.8A reactivity, confirming the specificity of this reagent. MoAb 5.8A was then used to examine the expression of MyoD1 in normal and malignant human tissues. MyoD1 was not detected in any normal adult tissue but was detected in 25 of 25 histologically confirmed rhabdomyosarcomas. Staining was localized to the nucleus and showed marked heterogeneity between cells as well as differential staining within nuclei. Specific subcellular localization of 5.8A was further determined by immunoelectron microscopy, where antibody was found to localize to electron-dense areas, more frequently associated with the nuclear submembranous region. In addition to rhabdomyosarcomas, MoAb 5.8A stained 2 of 5 Wilms9 tumors and one ectomesenchymoma, neoplasms known to contain myogenic elements. The 5.8A reagent was also of value in the accurate histopathological classification of 2 of 4 tumors previously diagnosed as extraosseous Ewing9s sarcoma and 2 of 3 tumors diaonnsed as undifferenriated sarCOmas. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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MyoDl Protein Expression in Alveolar Soft Part Sarcoma As Confirmatory Evidence of Its Skeletal Muscle Nature
The American journal of surgical pathology, 1991Co-Authors: Juan Rosai, David M. Parham, David N. Shapiro, Peter Dias, Peter J. HoughtonAbstract:A typical case of alveolar soft part sarcoma was found to express in a strong and widespread fashion the marker MyoD1 Protein. This nuclear phosphoProtein is the product of a regulatory gene that controls the commitment of a cell to myogenic lineage; it therefore provides strong and perhaps definitive evidence in support of the skeletal muscle nature of this enigmatic neoplasm.
P Dias - One of the best experts on this subject based on the ideXlab platform.
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Monoclonal antibodies to the myogenic regulatory Protein MyoD1: epitope mapping and diagnostic utility.
Cancer research, 1992Co-Authors: P Dias, David M. Parham, David N. Shapiro, Stephen J. Tapscott, Peter J. HoughtonAbstract:Monoclonal antibodies (MoAbs) were developed against recombinant wild-type murine MyoD1 Protein. Each of 4 MoAbs was immunologically reactive with recombinant MyoD1 Protein by enzyme-linked immunosorbent assay, and each specifically stained the nuclei of myogenic cells. Epitopes were mapped using fusion Protein constructs with specific deletions of defined regions of the MyoD1 molecule. MoAb 5.2F recognized an epitope in the amino terminal region between amino acid residues (AAR) 3 and 56, whereas epitopes for MoAbs 1.1A, 5.4G, and 5.8A were in the carboxyl terminus (AAR 167-318) of the MyoD1 Protein. The epitope for MoAb 5.8A was further delineated to AAR 170-209 by Western analysis and immunoprecipitation of in vivo transcribed and translated MyoD1 Protein having specific deletions in the carboxyl terminus. The 5.8A epitope was ultimately localized to the region between AAR 180 and 189 of the Protein by enzyme-linked immunosorbent assay using 10-amino acid residue synthetic peptides. This sequence is apparently unique to MyoD1 and has little homology to other myogenic regulatory Proteins (myogenin, Myf5, Myf6, and MRF4). Transfection of cDNA for murine MyoD1 into a nonmuscle cell line conferred 5.8A reactivity, confirming the specificity of this reagent. MoAb 5.8A was then used to examine the expression of MyoD1 in normal and malignant human tissues. MyoD1 was not detected in any normal adult tissue but was detected in 25 of 25 histologically confirmed rhabdomyosarcomas. Staining was localized to the nucleus and showed marked heterogeneity between cells as well as differential staining within nuclei. Specific subcellular localization of 5.8A was further determined by immunoelectron microscopy, where antibody was found to localize to electron-dense areas, more frequently associated with the nuclear submembranous region. In addition to rhabdomyosarcomas, MoAb 5.8A stained 2 of 5 Wilms' tumors and one ectomesenchymoma, neoplasms known to contain myogenic elements. The 5.8A reagent was also of value in the accurate histopathological classification of 2 of 4 tumors previously diagnosed as extraosseous Ewing's sarcoma and 2 of 3 tumors diagnosed as undifferentiated sarcomas.
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monoclonal antibodies to the myogenic regulatory Protein MyoD1 epitope mapping and diagnostic utility
Cancer Research, 1992Co-Authors: P Dias, David M. Parham, David N. Shapiro, Stephen J. Tapscott, Peter J. HoughtonAbstract:Abstract Monoclonal antibodies (MoAbs) were developed against recombinant wild-type murine MyoD1 Protein. Each of 4 MoAbs was immunologically reactive with recombinant MyoD1 Protein by enzyme-linked immunosorbent assay, and each specifically stained the nuclei of myogenic cells. Epitopes were mapped using fusion Protein constructs with specific deletions of defined regions of the MyoD1 molecule. MoAb 5.2F recognized an epitope in the amino terminal region between amino acid residues (AAR) 3 and 56, whereas epitopes for MoAbs 1.1A, 5.4G, and 5.8A were in the carboxyl terminus (AAR 167–318) of the MyoD1 Protein. The epitope for MoAb 5.8A was further delineated to AAR 170–209 by Western analysis and immunoprecipitation of in vitro transcribed and translated MyoDl Protein having specific deletions in the carboxyl terminus. The 5.8A epitope was ultimately localized to the region between AAR 180 and 189 of the Protein by enzyme-linked immunosorbent assay using 10-amino acid residue synthetic peptides. This sequence is apparently unique to MyoD1 and has little homology to other myogenic regulatory Proteins (myogenin, Myf5, Myf6, and MRF4). Transfection of cDNA for murine MyoDl into a nonmuscle cell line conferred 5.8A reactivity, confirming the specificity of this reagent. MoAb 5.8A was then used to examine the expression of MyoD1 in normal and malignant human tissues. MyoD1 was not detected in any normal adult tissue but was detected in 25 of 25 histologically confirmed rhabdomyosarcomas. Staining was localized to the nucleus and showed marked heterogeneity between cells as well as differential staining within nuclei. Specific subcellular localization of 5.8A was further determined by immunoelectron microscopy, where antibody was found to localize to electron-dense areas, more frequently associated with the nuclear submembranous region. In addition to rhabdomyosarcomas, MoAb 5.8A stained 2 of 5 Wilms9 tumors and one ectomesenchymoma, neoplasms known to contain myogenic elements. The 5.8A reagent was also of value in the accurate histopathological classification of 2 of 4 tumors previously diagnosed as extraosseous Ewing9s sarcoma and 2 of 3 tumors diaonnsed as undifferenriated sarCOmas. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
Stephen J. Tapscott - One of the best experts on this subject based on the ideXlab platform.
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Monoclonal antibodies to the myogenic regulatory Protein MyoD1: epitope mapping and diagnostic utility.
Cancer research, 1992Co-Authors: P Dias, David M. Parham, David N. Shapiro, Stephen J. Tapscott, Peter J. HoughtonAbstract:Monoclonal antibodies (MoAbs) were developed against recombinant wild-type murine MyoD1 Protein. Each of 4 MoAbs was immunologically reactive with recombinant MyoD1 Protein by enzyme-linked immunosorbent assay, and each specifically stained the nuclei of myogenic cells. Epitopes were mapped using fusion Protein constructs with specific deletions of defined regions of the MyoD1 molecule. MoAb 5.2F recognized an epitope in the amino terminal region between amino acid residues (AAR) 3 and 56, whereas epitopes for MoAbs 1.1A, 5.4G, and 5.8A were in the carboxyl terminus (AAR 167-318) of the MyoD1 Protein. The epitope for MoAb 5.8A was further delineated to AAR 170-209 by Western analysis and immunoprecipitation of in vivo transcribed and translated MyoD1 Protein having specific deletions in the carboxyl terminus. The 5.8A epitope was ultimately localized to the region between AAR 180 and 189 of the Protein by enzyme-linked immunosorbent assay using 10-amino acid residue synthetic peptides. This sequence is apparently unique to MyoD1 and has little homology to other myogenic regulatory Proteins (myogenin, Myf5, Myf6, and MRF4). Transfection of cDNA for murine MyoD1 into a nonmuscle cell line conferred 5.8A reactivity, confirming the specificity of this reagent. MoAb 5.8A was then used to examine the expression of MyoD1 in normal and malignant human tissues. MyoD1 was not detected in any normal adult tissue but was detected in 25 of 25 histologically confirmed rhabdomyosarcomas. Staining was localized to the nucleus and showed marked heterogeneity between cells as well as differential staining within nuclei. Specific subcellular localization of 5.8A was further determined by immunoelectron microscopy, where antibody was found to localize to electron-dense areas, more frequently associated with the nuclear submembranous region. In addition to rhabdomyosarcomas, MoAb 5.8A stained 2 of 5 Wilms' tumors and one ectomesenchymoma, neoplasms known to contain myogenic elements. The 5.8A reagent was also of value in the accurate histopathological classification of 2 of 4 tumors previously diagnosed as extraosseous Ewing's sarcoma and 2 of 3 tumors diagnosed as undifferentiated sarcomas.
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monoclonal antibodies to the myogenic regulatory Protein MyoD1 epitope mapping and diagnostic utility
Cancer Research, 1992Co-Authors: P Dias, David M. Parham, David N. Shapiro, Stephen J. Tapscott, Peter J. HoughtonAbstract:Abstract Monoclonal antibodies (MoAbs) were developed against recombinant wild-type murine MyoD1 Protein. Each of 4 MoAbs was immunologically reactive with recombinant MyoD1 Protein by enzyme-linked immunosorbent assay, and each specifically stained the nuclei of myogenic cells. Epitopes were mapped using fusion Protein constructs with specific deletions of defined regions of the MyoD1 molecule. MoAb 5.2F recognized an epitope in the amino terminal region between amino acid residues (AAR) 3 and 56, whereas epitopes for MoAbs 1.1A, 5.4G, and 5.8A were in the carboxyl terminus (AAR 167–318) of the MyoD1 Protein. The epitope for MoAb 5.8A was further delineated to AAR 170–209 by Western analysis and immunoprecipitation of in vitro transcribed and translated MyoDl Protein having specific deletions in the carboxyl terminus. The 5.8A epitope was ultimately localized to the region between AAR 180 and 189 of the Protein by enzyme-linked immunosorbent assay using 10-amino acid residue synthetic peptides. This sequence is apparently unique to MyoD1 and has little homology to other myogenic regulatory Proteins (myogenin, Myf5, Myf6, and MRF4). Transfection of cDNA for murine MyoDl into a nonmuscle cell line conferred 5.8A reactivity, confirming the specificity of this reagent. MoAb 5.8A was then used to examine the expression of MyoD1 in normal and malignant human tissues. MyoD1 was not detected in any normal adult tissue but was detected in 25 of 25 histologically confirmed rhabdomyosarcomas. Staining was localized to the nucleus and showed marked heterogeneity between cells as well as differential staining within nuclei. Specific subcellular localization of 5.8A was further determined by immunoelectron microscopy, where antibody was found to localize to electron-dense areas, more frequently associated with the nuclear submembranous region. In addition to rhabdomyosarcomas, MoAb 5.8A stained 2 of 5 Wilms9 tumors and one ectomesenchymoma, neoplasms known to contain myogenic elements. The 5.8A reagent was also of value in the accurate histopathological classification of 2 of 4 tumors previously diagnosed as extraosseous Ewing9s sarcoma and 2 of 3 tumors diaonnsed as undifferenriated sarCOmas. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
