Skip to main content
PMC home page
Plant Physiology logoLink to Plant Physiology
. 1987 Nov;85(3):835–840. doi: 10.1104/pp.85.3.835

Monoclonal Antibodies Against Fusicoccin with Binding Characteristics Similar to the Putative Fusicoccin Receptor of Higher Plants 1

Martin Feyerabend 1, Elmar W Weiler 1
PMCID: PMC1054348  PMID: 16665786

Abstract

Monoclonal antibodies were raised against fusicoccin. The toxin, linked to bovine serum albumin through its t-pentenyl moiety, served as immunogen. Hybridomas secreting anti-fusicoccin antibodies were screened by radioimmunoassay employing a novel radioactive derivative, [3H]-nor-fusicoccin-alcohol of high specific activity (1.5 × 1014Bq/mole). The two monoclonal antibodies reported here are of high apparent affinity for fusicoccin (0.71 × 10−9 molar and 1.85 × 10−9 molar). This is comparable to the apparent affinity of rabbit antiserum raised against the same type of conjugate (9.3 × 10−9 molar). A method for the single step purification of the monoclonal antibodies from ascites fluid is reported. A solid-phase immunoassay, using alkaline phosphatase as enzyme, exhibits a measuring range from 0.1 to 1.5 picomoles (about 70 picograms to 1 nanogram) of fusicoccin. The displacement of [3H]-nor-fusicoccin-alcohol from the antibodies by compounds structurally related to fusicoccin exhibits similar selectivity as a microsomal binding assay with the same tracer as radiolabeled probe.

Full text

PDF
835

Images in this article

837Fig. 2
on p.837

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  2. Burchiel S. W., Billman J. R., Alber T. R. Rapid and efficient purification of mouse monoclonal antibodies from ascites fluid using high performance liquid chromatography. J Immunol Methods. 1984 Apr 13;69(1):33–42. doi: 10.1016/0022-1759(84)90274-6. [DOI] [PubMed] [Google Scholar]
  3. Deschamps J. R., Hildreth J. E., Derr D., August J. T. A high-performance liquid chromatographic procedure for the purification of mouse monoclonal antibodies. Anal Biochem. 1985 Jun;147(2):451–454. doi: 10.1016/0003-2697(85)90296-9. [DOI] [PubMed] [Google Scholar]
  4. Eberle J., Arnscheidt A., Klix D., Weiler E. W. Monoclonal Antibodies to Plant Growth Regulators: III. Zeatinriboside and Dihydrozeatinriboside. Plant Physiol. 1986 Jun;81(2):516–521. doi: 10.1104/pp.81.2.516. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kearney J. F., Radbruch A., Liesegang B., Rajewsky K. A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immunol. 1979 Oct;123(4):1548–1550. [PubMed] [Google Scholar]
  6. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  7. OUCHTERLONY O. Diffusion-in-gel methods for immunological analysis. Prog Allergy. 1958;5:1–78. [PubMed] [Google Scholar]
  8. Rasi-Caldogno F., Pugliarello M. C. Fusicoccin stimulates the H+-ATPase of plasmalemma in isolated membrane vesicles from radish. Biochem Biophys Res Commun. 1985 Nov 27;133(1):280–285. doi: 10.1016/0006-291x(85)91872-8. [DOI] [PubMed] [Google Scholar]
  9. Stanker L. H., Vanderlaan M., Juarez-Salinas H. One-step purification of mouse monoclonal antibodies from ascites fluid by hydroxylapatite chromatography. J Immunol Methods. 1985 Jan 21;76(1):157–169. doi: 10.1016/0022-1759(85)90488-0. [DOI] [PubMed] [Google Scholar]
  10. Stout R. G., Cleland R. E. Partial characterization of fusicoccin binding to receptor sites on oat root membranes. Plant Physiol. 1980 Sep;66(3):353–359. doi: 10.1104/pp.66.3.353. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES