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Histochemistry

, Volume 50, Issue 4, pp 271–279 | Cite as

Production of specific antibodies to contractile proteins, and their use in immunofluorescence microscopy

II. Species-specific and species-non-specific antibodies to smooth and striated chicken muscle actin
  • Ute Gröschel-Stewart
  • Susi Ceurremans
  • Irmtraut Lehr
  • Christine Mahlmeister
  • Eltriede Paar
Article

Summary

The injection of rabbits with insoluble or soluble G-actin from chicken smooth or striated muscle will produce antibodies that are equally reactive, and species and tissue non-specific in immunoprecipitation, immunofluorescence and actin-activated Mg2+-ATPase** inhibition test. These antibodies have been used for the identification of actin-containing fibrils in a variety of tissues. When G-actins from chicken smooth or striated muscle are immobilized by chemical linkage to Affi-Gel 702 microbeads, their immunogenicity is increased, but the antibodies obtained against them are species-specific and will only react with actin and actin-containing structures from chicken and are therefore limited in use. It is concluded from this work that insoluble G-actin is the preferable immunogen to obtain precipitating antibodies for wide use.

Keywords

Public Health Striate Muscle Fibril Specific Antibody Immunofluorescence Microscopy 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Abbreviations

ATPase

Adenosinetriphosphatase

FITC

Fluoresceinisothiocyanate

SDS

Sodiumdodecylsulfate

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References

  1. Amsterdam, A., Lamed, R., Josephs, R., Silman, I., Gröschel-Stewart, U.: Actomyosin from electric organ tissue of the electric eel, Electrophorus electricus. Abstracts, Sixth European Congress on Electron Microscopy, Jerusalem, September 1976Google Scholar
  2. Amsterdam, A., Lindner, H.R., Gröschel-Stewart, U.: Immunofluorescent localization of actin and myosin in the rat ovary. Abstracts 16. J. Cell Biol 67, 8a (1975)Google Scholar
  3. Anderson, P.J.: The actin of muscle and fibroblasts. Biochem. J. 155, 297–301 (1976)Google Scholar
  4. Campbell, D.H., Garvey, J.S., Cremer, N.E., Sussdorf, D.H.: In: Methods in immunology, p. 118. New York: W.A. Benjamin 1964Google Scholar
  5. Carsten, E., Mommaerts, W.F.H.M.: A study of actin by means of starch gel electrophoresis. Biochemistry, (Wash.) 2, 28–32 (1963)Google Scholar
  6. Chamley, J.H., Gröschel-Stewart, U., Campbell, G.R.: Distinction between dedifferentiated smooth muscle cells and fibroblasts in culture. Abstracts, International Symposium on Contractile Systems in Non-muscle Cells. Bressanone, September 1976Google Scholar
  7. Elzinga, M., Collins, J.H.: The amino acid sequence of rabbit skeletal muscle actin. In: The mechanism of muscle contraction, Vol XXXVII. Cold Spring Harbor Symposia on Quantitative Biology, p. 1–7. Cold Spring Harbor Laboratory 1973Google Scholar
  8. Elzinga, M., Maron, B.J., Adelstein, R.S.: Human heart and platelet actins are products of different genes. Science 191, 94–95 (1976)Google Scholar
  9. Gabbiani, G., Ryan, G.B., Lamelin, J.-P., Vassalli, P., Majno, G., Bouvier, C.A., Cruchaud, A., Lüscher, E.F.: Human smooth muscle autoantibody. Amer. J. Path. 72, 473–484 (1973)Google Scholar
  10. Gosselin-Rey, C., Gerday, Ch., Gaspar-Godfroid, A., Carsten, M.E.: Amino acid analysis and peptide mapping of bovine carotid actin. Biochim. biophys. Acta (Amst.) 175, 165–173 (1969)Google Scholar
  11. Gröschel-Stewart, U., Chamley, J.H., McConnell, J.D., Burnstock, G.: Comparison of the reaction of cultured smooth and cardiac muscle cells and fibroblasts to specific antibodies to myosin. Histochemistry 43, 215–224 (1975)Google Scholar
  12. Gröschel-Stewart, U., Schreiber, J., Mahlmeister, Ch., Weber, K.: Production of specific antibodies to contractile proteins, and their use in immunofluorescence microscopy. I. Antibodies to smooth and striated chicken muscle myosins. Histochemistry 46, 229–236 (1976)Google Scholar
  13. Gruenstein, E., Rich, A.: Non-identity of muscle and non-muscle actins. Biochem. biophys. Res. Commun. 64 472–477 (1975)Google Scholar
  14. Heine, W.D., Altmann, H.W., Gröschel-Stewart, U.: Aktin und Myosin in Leberepithelien. Abstracts, Deutscher Pathologen-Kongress, Freiburg, Juni 1976Google Scholar
  15. Hirabayashi, T., Hayashi, Y.: Antibody specific for action from frog skeletal muscle. J. Biochem. (Tokyo) 71, 153–156 (1972)Google Scholar
  16. Humphrey, J.H., White, R.G.: Kurzes Lehrbuch der Immunologie, p. 133, ed. E. Macher. Stuttgart: G. Thieme 1972Google Scholar
  17. Lazarides, E., Weber, K.: Actin antibody: The specific visualization of actin filaments in non-muscle cells. Proc. nat. Acad. Sci. (Wash.) 71, 2268–2272 (1974)Google Scholar
  18. Lidman, K., Biberfeld, G., Fagraeus, A., Norberg, R., Torstensson, R., Utter, G., Carlsson, L., Luca, J., Lindberg, U.: Anti-actin specificity of human smooth muscle antibodies in chronic active hepatitis. Clin. exp. Immunol. 24, 266–272 (1976)Google Scholar
  19. Owaribe, K., Hatano, S.: Induction of antibody against actin from myxomycete plasmodium and its properties. Biochemistry (Wash.) 14, 3024–3029 (1975)Google Scholar
  20. Spudich, J.A., Watt, S.: The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin. J. biol. Chem. 246, 4866–4871 (1971)Google Scholar
  21. Storti, R.V., Rich, A.: Chick cytoplasmic actin and muscle actin have different structural genes. Proc. nat. Acad. Sci. (Wash.) 73, 2346–2350 (1976)Google Scholar
  22. Weber, K., Gröschel-Stewart, U.: Antibody to myosin: The specific visualization of myosin-containing filaments in nonmuscle cells. Proc. nat. Acad. Sci. (Wash.) 71, 4561–4564 (1974)Google Scholar
  23. Weber, K., Osborn, M.: The reliability of molecular weight determinations by dodecyl-sulfatepolyacrylamide gel electrophoresis. J. biol. Chem. 244, 4406–4412 (1969)Google Scholar

Copyright information

© Springer-Verlag 1977

Authors and Affiliations

  • Ute Gröschel-Stewart
    • 1
  • Susi Ceurremans
    • 1
  • Irmtraut Lehr
    • 1
  • Christine Mahlmeister
    • 1
  • Eltriede Paar
    • 1
  1. 1.Institut für Zoologie der Technischen Hochschule DarmstadtDarmstadtGermany

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