title: Production of a rat monoclonal antibody specific for Myf5.
abstract: Myogenic regulatory factors (MRFs) are transcription factors that possess a characteristic basic helix-loop-helix domain. Myf5, MyoD, MRF4, and myogenin are well-known MRF family members that activate muscle-specific genes during differentiation. Myf5 is expressed first among MRFs at the very early phase and plays an important role in myoblast specificity and cell proliferation. Myf5 shares high homology with MyoD, and therefore some commercial Myf5 antibodies are cross-reactive for Myf5 and MyoD. To allow for detailed studies of the function of Myf5, we generated a monoclonal antibody specific for Myf5 utilizing a rat medial iliac lymph node method. Immunoblot analysis using our monoclonal antibody enabled us to identify Myf5 protein from rat myoblast L6E9 cell extract. Moreover, cell immunostaining revealed the nuclear localization of Myf5 in the L6E9 cells. This monoclonal antibody against Myf5 will allow us to perform further detailed studies of Myf5 and Myf5 function.
authors.affiliation: SSP Stem Cell Unit, Faculty of Medicine, Kyushu University , Fukuoka, Japan .
authors.forename: Akihito
authors.initials: A
authors.lastname: Harada
authors.forename: Seiji
authors.initials: S
authors.lastname: Okada
authors.forename: Jun
authors.initials: J
authors.lastname: Odawara
authors.forename: Hiromi
authors.initials: H
authors.lastname: Kumamaru
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Saiwai
authors.forename: Mayumi
authors.initials: M
authors.lastname: Aoki
authors.forename: Mako
authors.initials: M
authors.lastname: Nakamura
authors.forename: Yuko
authors.initials: Y
authors.lastname: Nishiyama
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Ohkawa
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology* / Q000302:isolation & purification; D000918:Antibody Specificity; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006825:Hybridomas / Q000378:metabolism; D007082:Ileum / Q000276:immunology; D007114:Immunization; D007124:Immunoenzyme Techniques; D008198:Lymph Nodes / Q000166:cytology / Q000276:immunology; D051379:Mice; D032446:Myoblasts / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D051787:Myogenic Regulatory Factor 5 / Q000235:genetics / Q000276:immunology*; D041681:NIH 3T3 Cells; D010957:Plasmids; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; C497418:Myf5 protein, mouse; D051787:Myogenic Regulatory Factor 5; D011994:Recombinant Proteins