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Enzyme-Linked Immunosorbent Assay

Enzyme-Linked Immunosorbent Assay

In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or quantified.

Enzyme-linked Receptors

Enzyme-linked Receptors

Enzyme-linked receptors are proteins that act as both receptor and enzyme, activating multiple intracellular signals. This is a large group of receptors that include the receptor tyrosine kinase (RTK) family. Many growth factors and hormones bind to and activate the RTKs.
Neurotrophin (NT) receptors are a family of RTKs, including trkA, trkB, and trkC (tropomyosin-related kinase) receptors. TrkA is specific for nerve growth factor (NGF), neurotrophin-6, and neurotrophin-7. TrkB binds...

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Related Experiment Video

Updated: Jul 5, 2026

Characterization of Thymus-dependent and Thymus-independent Immunoglobulin Isotype Responses in Mice Using Enzyme-linked Immunosorbent Assay

Characterization of Thymus-dependent and Thymus-independent Immunoglobulin Isotype Responses in Mice Using Enzyme-linked Immunosorbent Assay

Published on: September 7, 2018

Enzyme-linked immunosorbent assays.

¹University of Maryland, Baltimore, Maryland, USA.

Current Protocols in Immunology

|April 25, 2008

Summary

This summary is machine-generated.

This study details six enzyme-linked immunosorbent assay (ELISA) systems for detecting antibodies and antigens. These methods utilize enzyme-conjugated reagents and substrate hydrolysis to quantify analytes, enabling visual or instrument-based detection.

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Area of Science:

Immunology
Biochemistry
Analytical Chemistry

Background:

Enzyme-linked immunosorbent assays (ELISA) are widely used for detecting specific antibodies and antigens.
Standard ELISA protocols involve immobilizing reactants on solid phases for detection.

Purpose of the Study:

To describe six distinct ELISA systems for detecting specific antibodies, soluble antigens, or cell-surface antigens.
To provide methods for optimizing ELISA protocols and preparing enzyme conjugates.

Main Methods:

Six ELISA systems are presented, utilizing solid-phase reactants (adsorbed antigens/antibodies or cell-associated molecules).
Soluble reactants bind to solid-phase reactants, followed by incubation with enzyme-conjugated secondary or tertiary reactants.
Detection involves adding a substrate that is hydrolyzed by the bound enzyme to produce a detectable signal (colorimetric or fluorometric).

Main Results:

The amount of generated product is directly proportional to the concentration of the analyte.
The described protocols allow for the detection of various analytes, including antibodies and cell-surface antigens.
A support protocol for optimizing ELISAs and preparing alkaline phosphatase conjugates is included.

Conclusions:

The presented ELISA systems offer versatile and sensitive methods for analyte detection.
These protocols can be optimized for specific applications, enhancing their utility in biological and medical research.
The ability to detect various antigens and antibodies makes these ELISAs valuable tools in diagnostics and research.