title: Generation of monoclonal antibodies against chromosomal antigens that have a high sequence similarity between human and mouse.
abstract: We raised monoclonal antibodies by immunizing mice with total chromosome proteins extracted from isolated human metaphase chromosomes. The indirect immunofluorescence screening of hybridoma cell lines provided 15 monoclonal antibodies against the chromosomal antigens. The antigen proteins of the mAbs were identified by immunoblotting as core histones or by immunoprecipitation followed by a peptide mass fingerprinting method as nuclear mitotic apparatus protein, heterogeneous nuclear ribonucleoprotein A2/B1, ribosomal protein S4, linker histone and beta-actin. During mitosis, localizations of these proteins on chromosomes were clearly observed using the obtained antibodies. These results indicate that the current strategy is effective for obtaining monoclonal antibodies useful for immunoblotting and/or immunofluorescent staining of human proteins, using the antigens with high homology to mouse proteins.
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Tsunehito
authors.initials: T
authors.lastname: Higashi
authors.forename: Shuichi
authors.initials: S
authors.lastname: Miyakawa
authors.forename: Susumu
authors.initials: S
authors.lastname: Uchiyama
authors.forename: Sachihiro
authors.initials: S
authors.lastname: Matsunaga
authors.forename: Hideaki
authors.initials: H
authors.lastname: Takata
authors.forename: Satoru
authors.initials: S
authors.lastname: Fujimoto
authors.forename: Masanori
authors.initials: M
authors.lastname: Noda
authors.forename: Akiko
authors.initials: A
authors.lastname: Terauchi
authors.forename: Takeyuki
authors.initials: T
authors.lastname: Shimizu
authors.forename: Masayuki
authors.initials: M
authors.lastname: Oda
authors.forename: Takachika
authors.initials: T
authors.lastname: Azuma
authors.forename: Kiichi
authors.initials: K
authors.lastname: Fukui
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology*; D000918:Antibody Specificity; D000941:Antigens / Q000276:immunology*; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis* / Q000276:immunology; D002877:Chromosomes, Human / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D019084:Fluorescent Antibody Technique, Indirect; D006367:HeLa Cells; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007116:Immunization, Passive; D015151:Immunoblotting; D008214:Lymphocytes / Q000166:cytology / Q000276:immunology; D013058:Mass Spectrometry; D008677:Metaphase; D051379:Mice; D009101:Multiple Myeloma / Q000276:immunology / Q000473:pathology; D010449:Peptide Mapping; D011233:Precipitin Tests
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens; D002868:Chromosomal Proteins, Non-Histone