Use of monoclonal antibody to detect bone morphogenetic protein-4 (BMP-4).
A monoclonal antibody that reacts with murine and human bone morphogenetic protein-4 (BMP-4) has been developed using recombinant BMP-4 as an immunogen. The antibody that bound most tightly to recombinant murine (rm)BMP-4 was selected, subcloned, and characterized. The specificity of the antibody was confirmed using Western blot analysis and enzyme-linked immunosorbent assay (ELISA). The antibody reacts with murine and human BMP-4 in both the reduced and nonreduced condition; however, this antibody shows cross-reactivity with neither human BMP-2 nor TGF-beta 1. Thus, the produced antibody could recognize the disulfide-linked dimeric structure of bioactive BMP-4, regardless of the species. Immunocytochemical study using this antibody successfully shows the cytosolic localization of BMP-4 in osteoinductive cells; i.e., BFO and Saos-2 in which the level of mRNA for BMP-4 was proved to be constitutively high by Northern blot analysis. In addition, the antibody could demonstrate the presence of BMP-4 in developmental bone formation in the alveolar bone of rat embryo by immunohistochemistry. The antibody could be used for a more sensitive approach for quantitative analysis of BMP-4.
Masuhara
K
K
Department of Orthopaedic Surgery, Osaka University Medical School, Suita, Japan.
Nakase
T
T
Suzuki
S
S
Takaoka
K
K
Matsui
M
M
Anderson
H C
HC
D000818:Animals; D000911:Antibodies, Monoclonal; D019485:Bone Morphogenetic Proteins; D002460:Cell Line; D005314:Embryonic and Fetal Development / Q000502:physiology; D004926:Escherichia coli; D005326:Fetal Proteins / Q000032:analysis; D006801:Humans; D051379:Mice; D009363:Neoplasm Proteins / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D014176:Protein Biosynthesis; D011506:Proteins / Q000032:analysis* / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis
D000911:Antibodies, Monoclonal; D019485:Bone Morphogenetic Proteins; D005326:Fetal Proteins; D009363:Neoplasm Proteins; D011506:Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins