Summary
Monoclonal and polyclonal antibodies were raised against 150k, 60k, 32k, and 15k phosphoprotein components of 14-week chicken bone. Hybridomas were prepared by immunizing Balb/c mice with 150k, 60k, or 32k phosphoproteins followed by fusion of their spleen cells with X63-Ag8.653 myeloma cells. Polyclonal antibodies to the 60k, 32k, and 15k phosphoprotein components were produced in immunized rabbits. Immunological cross-reactivity between antigen and antibody was examined by enzyme-linked immunosorbent assay (ELISA) and dot-blotting. There was cross-reactivity between the 150k, 32k, and 15k phosphoprotein components and between the 150k, 90K, and 60k components. The antibodies raised against 60k component do not bind 32k and 15k antigens.
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References
Glimcher MJ (1984) Recent studies of the mineral phase in bone and its possible linkage to the organic matrix by protein-bound phosphate bonds. Phil Trans Roy Soc 340(B):479–508
Veis A, Stetler-Stevenson W, Takagi Y, Sabsay B, Fullerton R (1981) The nature and localization of phosphorylated proteins of mineralized dentin. In: Veis A (ed) The chemistry and biology of mineralized connective tissues. Elsevier/North-Holland, New York, pp 377–387
Uchiyama A, Suzuki M, Lefteriou B, Glimcher MJ (1986) Isolation and chemical characterization of the phosphoproteins of chicken bone matrix: heterogeneity in molecular weight and composition. Biochem 25:7572–7583
Lee SL, Kossiva D, Glimcher MJ (1983) Phosphoproteins of bovine dentin: evidence for polydispersity during tooth maturation. Biochem 22:2596–2601
Dimuzio MT, Bhown M, Butler WT (1985) The biosynthesis of dentin phosphophoryns by rat incisor odontoblasts in organ culture. Calcif Tissue Int 37:242–249
Masters PM (1985) In vivo decomposition of phosphoserine and serine in noncollagenous protein from human dentin. Calcif Tissue Int 37:236–241
Tsay TG, Veis A (1985) The preparation, detection and characterization of an antibody to rat phosphophoryn. Biochem 24:6363–6369
Engvall E, Perlmann P (1972) Enzyme-linked immunoabsorbent assay, III. Quantification of specific antibodies of enzyme-labeled anti-immunoglobulin in antigen coated tubes. J Immunol 109:129–135
Kearney JF, Radbruch A, Liesegang B, Rajewsky K (1979) A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immunol 123:1548–1550
Littlefield JW (1964) Selection of hybrids from matings of fibroblasts in vitro and their presumed recombinants. Science 145:709–710
Hawkes R, Niday E, Gordon J (1982) A dot-immunobinding assay for monoclonal and other antibodies. Anal Biochem 119:142–147
Franzen A, Heinegard D (1985) The chemistry and biology of mineralized tissues. Butler WT (ed) EBSCO Media, Inc. Birmingham, Alabama, pp 132–141
Prince CW, Oosawa T, Butler WT, Tomana M, Bhown AS, Bhown M, Schrohenloher RE (1987) Isolation characterization and biosynthesis of a phosphorylated protein from rat bone. J Biol Chem 262:2900–2907
Oldberg A, Franzen A, Heinegard D (1986) Cloning and sequence analysis of rat bone sialoprotein (osteopontin) cDNA reveals an Arg-Gly-Asp cell-binding sequence. Proc Natl Acad Sci 83:8819–8823
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Yamazaki, K., Suzuki, M., Mikuni-Takagaki, Y. et al. Preparation of monoclonal antibodies to chicken bone phosphoproteins. Calcif Tissue Int 43, 41–43 (1988). https://doi.org/10.1007/BF02555166
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DOI: https://doi.org/10.1007/BF02555166