Advertisement

An Enzyme-Linked Immunosorbent Assay for Calmodulin

  • M. E. Bardsley
  • P. J. Roberts
  • I. R. Cottingham
Part of the Methodological Surveys in Biochemistry and Analysis book series (MSBA, volume 15B)

Abstract

Calmodulin (CaM) is a Ca2+-binding protein notably important in the translation of Ca2+ signals inside the cell (as reviewed [1, 2]) [cf. A.R. Means & J.G. Chafouleas, Vol. 13, #A-7, this series — Ed.]. It is found throughout the plant and animal kingdoms and its sequence is highly conserved [3]. Measurement of CaM by its activation of 3’,5’-cyclic nucleotide phosphodiesterase, and using radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA), provides useful comparative measurements of the amount of protein and its activity. Methods have been described for assay by enzyme activation [4] and by RIA ([5, 6], & Vol. 13 art. cited above). Here we describe the production of an anti-CaM antibody (Ab) and its use in setting up an ELISA. Elsewhere [7] we exemplify the applicability of the ELISA.

Keywords

Ammonium Sulphate Cyclic Nucleotide Phosphodiesterase DEAE Column Polystyrene Cuvette Cuvette Wall 
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.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Klee, C.B., Crouch, T.H. & Richmann, P.G. (1980) Ann. Rev. Biochem. 49, 489–515.Google Scholar
  2. 2.
    Kakiuchi, S., Hidaka, H. & Means, A.R., eds. (1982) Calmodulin and Intracellular Calcium Receptors, Plenum, New York, 425 pp.CrossRefGoogle Scholar
  3. 3.
    Vanaman, T.C. (1980) in Calcium and Cell Function, Vol. 1 (Cheung, W.Y., ed.), Academic Press, New York, PP. 41–58.Google Scholar
  4. 4.
    Wallace, R.W., Tallant, E.A. & Cheung, W.Y. (1983) Meths. Enzymol. 102, 39–47.CrossRefGoogle Scholar
  5. 5.
    Chafouleas, J.G., Dedman, J.R., Munjaal, R.P. & Means, A.R. (1979) J. Biol. Chem. 254, 10262–10267.Google Scholar
  6. 6.
    Wallace, R.W. & Cheung, W.Y. (1979) J. Biol. Chem. 254, 6564–6571.Google Scholar
  7. 7.
    Bardsley, M.E., Roberts, P.J. & Cottingham, I.R. (1985) Biochem. Soc. Trans., in press.Google Scholar
  8. 8.
    Kakiuchi, S., Sobue, K., Yamazaki, R., Kambayashi, J., Sakon, M. & Kosaki, G. (1981) FEBS Lett. 126, 203–207.CrossRefGoogle Scholar
  9. 9.
    Hirs, C.H.W. (1967) Meths. Enzymol. 11, 197–199.CrossRefGoogle Scholar
  10. 10.
    Van Eldik, L.J. & Watterson, D.M. (1981) J. Biol. Chem. 256, 4205–4210.Google Scholar
  11. 11.
    Hurn, B.A.L. & Chantler, S.M. (1980) Meths. Enzymol. 70, 104–142Google Scholar
  12. 12.
    Engvall, E. & Perlmann, P. (1972) J. Immunol. 109, 129–135.Google Scholar

Copyright information

© Plenum Press, New York 1985

Authors and Affiliations

  • M. E. Bardsley
    • 1
  • P. J. Roberts
    • 1
  • I. R. Cottingham
    • 2
  1. 1.Department of Physiology and Pharmacology Medical & Biological SciencesUniversity of SouthamptonSouthamptonUK
  2. 2.Department of BiochemistryUniversity of SussexFalmer, BrightonUK

Personalised recommendations