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The Use of Enzyme-Linked Immunosorbent Assays to Quantitate Proteins in Biological Functions

  • Dori R. Germolec
Part of the Methods in Molecular Medicine™ book series (MIMM, volume 22)

Abstract

Immunoassays are techniques for measuring the concentration or activity of a substance using immunological reactions. Several types of immunoassay are commonly used, including precipitation assays using antibody/antigen complexes, agglutination assays using coated erythrocytes or other particles, and radio-(RIA) and enzyme immunoassays. Multiple factors determine the best assay for a particular use, including sample concentration, technical difficulty, required precision, and availability of specialized equipment. The most sensitive assays are the radio- and enzyme immunoassays, however safety and environmental issues regarding the use of radioisotopes have limited the application of RIAs, and thus enzyme-linked immunosorbent assays (ELISA) tend to be the method of choice when sample concentration is a critical issue. The potential for robotic automation and the availability of inexpensive microplate dilutors, pipetors, and readers has allowed for the generation of highly reproducible results for large numbers of samples in an short time.

Keywords

Capture Antibody Dilute Stock Solution Wash Plate Passive Adsorption Sandwich ELISAs 
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.

References

  1. 1.
    Rubenstein, K. E., Schneider, R. S., and Ullman, E. F. (1972) Homogeneous enzyme-immunoassay. A new immunochemical technique. Biochem. Biophys. Res. Commun. 47, 846–858.PubMedCrossRefGoogle Scholar
  2. 2.
    Ullman, E. F. and Maggio, E. T. (1980) Principles of homogeneous enzyme-immunoassay, in Enzyme-immunoassay (Maggio, E. T., ed.), CRC, Boca Raton, FL, pp. 106–131.Google Scholar
  3. 3.
    Crowther, J. R. (1995) ELISA: Theory and Practice. Humana, Totowa, NJ.Google Scholar
  4. 4.
    Pollock, P. L., Germolec, D. R., Comment, C. E., Rosenthal, G. J., and Luster, M. I. (1994) Human lymphocyte engrafted SCID mice as a model for immunotoxicity assessment: studies with cyclosporine A and TCDD. Fundam. Appl. Toxicol. 22, 130–138.PubMedCrossRefGoogle Scholar
  5. 5.
    Kemeny, D. M. (1990) A Practical Guide to ELISA. Pergamon, Oxford.Google Scholar
  6. 6.
    Carpenter, A. B. (1992). Enzyme-linked immunoassays, in Manual of Clinical Laboratory Immunology, 4th ed. (Rose, N. R., de Macario, E. C, Fahey, J. L., Friedman, H., and Penn, G. M., eds.), American Society for Microbiology, Washington, DC, pp. 2–9.Google Scholar
  7. 7.
    Macy, E., Kemeny, M., and Saxon, A. (1988) Enhanced ELISA: how to measure less than 10 picograms of a specific protein (immunoglobulin) in less than 8 hours. FASEB J. 2, 3003–3009.PubMedGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1999

Authors and Affiliations

  • Dori R. Germolec
    • 1
  1. 1.Laboratory of ToxicologyNational Institute of Environmental Health scienceResearch Triangle Park

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