Enzyme-linked immunosorbent assays

Current Protocols in Immunology
P Hornbeck

Abstract

This unit describes six different ELISA systems for the detection of specific antibodies, soluble antigens, or cell-surface antigens. In all six systems, soluble reactants are removed from solution after specifically binding to solid-phase reactants. In the first four protocols, solid-phase reactants are prepared by adsorbing an antigen or antibody onto plastic microtiter plates; in the next two protocols, the solid-phase reactants are cell-associated molecules. In all protocols, the solid-phase reagents are incubated with secondary or tertiary reactants covalently coupled to an enzyme. Unbound conjugates are washed out and a chromogenic or fluorogenic substrate is added. As the substrate is hydrolyzed by the bound enzyme conjugate, a colored or fluorescent product is generated. Finally, the product is detected visually or with a microtiter plate reader. The amount of product generated is proportional to the amount of analysate in the test mixture. The Support Protocol can be used to optimize the different ELISAs. The second support protocol presents a method for preparing alkaline phosphatase conjugates.

References

Feb 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·N HaradaA Miyajima
Jun 26, 1987·Journal of Immunological Methods·J D BeattyW G Vlahos
Dec 4, 1987·Journal of Immunological Methods·W C Bartlett, R J Noelle
Nov 1, 1988·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·E MacyA Saxon
May 9, 1988·Journal of Immunological Methods·A SchotsE Egberts
May 26, 1972·Biochemical and Biophysical Research Communications·K E RubensteinE F Ullman

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Citations

Jul 15, 2015·Bioanalysis·Cameron D YamanishiShuichi Takayama
Feb 24, 2009·Current Protocols in Microbiology·D'Ann Rochon
Jun 3, 2000·Carcinogenesis·B M SchmiedP M Pour
Apr 4, 2009·The Journal of Immunology : Official Journal of the American Association of Immunologists·Yvonne R ChanJay K Kolls
Sep 22, 2011·Current Protocols in Pharmacology·Matthew M SeaveyKristine L Stump
Jan 1, 2013·Current Protocols in Mouse Biology·Birgit RathkolbMartin Hrabě de Angelis
Jan 5, 2020·Cold Spring Harbor Protocols·Maya Z Piccinni, Matthew J Guille
Aug 21, 2020·BMC Biotechnology·Tania García-MaceiraElier Paz-Rojas
Jun 25, 2021·Expert Review of Clinical Immunology·Ali NiliMaryam Daneshpazhooh

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