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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
12
pubmed:dateCreated
1996-7-5
pubmed:abstractText
A monoclonal antibody (Th-10a) specific for the nuclear protein appearing in the S phase of the cell cycle in normal mouse thymocytes was derived by immunizing Wistar rats with a murine thymic lymphoma (TIGN), and its isotype was rat IgG2a and had kappa light chain. Immunohistochemical staining of frozen sections of B10.Thy1.1 newborn thymus and embryonic intestine revealed that this monoclonal antibody reacted strongly with the nuclear proteins of subcortical thymocytes and the basal layer of the mucosa, where many cells were dividing, but not with that of the thymic medullary area. To evaluate the expression of the nuclear proteins during the cell cycle in detail, the results of an immunofluorescence analysis of the thymocytes from hydroxyurea-treated B10 mice using Th-10a monoclonal antibody were compared with those of DNA synthesis of these cells with the use of the FITC-conjugated anti-BrdUrd monoclonal antibody. The results indicated that the nuclear protein detected by Th-10a monoclonal antibody was highly expressed in the S phase of normal thymocytes, while the cells in G1, G2 and M phases exhibited a low level of the expression. Moreover, the variations in expression of the nuclear proteins in the thymocytes at different times after hydroxyurea treatment were observed to correspond with the frequency of DNA synthesizing cells. In contrast, the high level and unregulated expression of the nuclear protein detected by Th-10a monoclonal antibody was observed throughout the cell cycle of the mouse lymphoma cell lines examined. Since Th-10a monoclonal antibody does not react with the nuclear proteins derived from human, hamster or rat proliferating cells, this antibody may recognize a murine specific epitope of the nuclear protein. To further characterize the nuclear proteins, we extracted them from normal thymocytes or thymic lymphomas, and analysed them by immunoblotting or immunoprecipitation followed by SDS-polyacrylamide gel electrophoresis. The nuclear protein(s) detected by Th-10a monoclonal antibody was mostly 95 kDa and also 83 kDa polypeptide. The results also indicated that the 95 kDa nuclear protein was phosphorylated in vivo.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0960-7722
pubmed:author
pubmed:issnType
Print
pubmed:volume
28
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
645-57
pubmed:dateRevised
2007-11-15
pubmed:meshHeading
pubmed-meshheading:8634372-Animals, pubmed-meshheading:8634372-Antibodies, Monoclonal, pubmed-meshheading:8634372-Cell Division, pubmed-meshheading:8634372-Cell Nucleus, pubmed-meshheading:8634372-Cricetinae, pubmed-meshheading:8634372-DNA, Neoplasm, pubmed-meshheading:8634372-DNA Replication, pubmed-meshheading:8634372-Female, pubmed-meshheading:8634372-Gene Expression Regulation, Neoplastic, pubmed-meshheading:8634372-Humans, pubmed-meshheading:8634372-Hydroxyurea, pubmed-meshheading:8634372-Immunoblotting, pubmed-meshheading:8634372-Immunoglobulin G, pubmed-meshheading:8634372-Lymphoma, T-Cell, pubmed-meshheading:8634372-Male, pubmed-meshheading:8634372-Mice, pubmed-meshheading:8634372-Microscopy, Fluorescence, pubmed-meshheading:8634372-Molecular Weight, pubmed-meshheading:8634372-Neoplasm Proteins, pubmed-meshheading:8634372-Neoplastic Stem Cells, pubmed-meshheading:8634372-Nuclear Proteins, pubmed-meshheading:8634372-Rats, pubmed-meshheading:8634372-Rats, Wistar, pubmed-meshheading:8634372-S Phase, pubmed-meshheading:8634372-T-Lymphocytes, pubmed-meshheading:8634372-Thymus Neoplasms
pubmed:year
1995
pubmed:articleTitle
The characterization of the monoclonal antibody Th-10a, specific for a nuclear protein appearing in the S phase of the cell cycle in normal thymocytes and its unregulated expression in lymphoma cell lines.
pubmed:affiliation
Division of Biology and Oncology, National Institute of Radiological Science, Chiba, Japan.
pubmed:publicationType
Journal Article