Production of a rat monoclonal antibody specific for Myf5.
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.
Harada
Akihito
A
SSP Stem Cell Unit, Faculty of Medicine, Kyushu University , Fukuoka, Japan .
Okada
Seiji
S
Odawara
Jun
J
Kumamaru
Hiromi
H
Saiwai
Hirokazu
H
Aoki
Mayumi
M
Nakamura
Mako
M
Nishiyama
Yuko
Y
Ohkawa
Yasuyuki
Y
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*
D000911:Antibodies, Monoclonal; C497418:Myf5 protein, mouse; D051787:Myogenic Regulatory Factor 5; D011994:Recombinant Proteins