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Related Concept Videos

Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

17.9K
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...
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Enzyme-linked Receptors01:00

Enzyme-linked Receptors

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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: Feb 27, 2026

An ELISA Based Binding and Competition Method to Rapidly Determine Ligand-receptor Interactions
08:40

An ELISA Based Binding and Competition Method to Rapidly Determine Ligand-receptor Interactions

Published on: March 14, 2016

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Direct Competitive Enzyme-Linked Immunosorbent Assay (ELISA).

Thomas O Kohl, Carl A Ascoli

    Cold Spring Harbor Protocols
    |July 7, 2017
    PubMed
    Summary

    The competitive enzyme-linked immunosorbent assay (ELISA) quantifies small-molecule antigens. Higher antigen concentrations in samples result in weaker signals, enabling accurate measurement in complex mixtures.

    Area of Science:

    • Immunochemistry
    • Analytical Chemistry

    Background:

    • The competitive enzyme-linked immunosorbent assay (cELISA) is a versatile immunoassay technique.
    • It relies on the principle of competition between a labeled analyte and the analyte in a sample for limited binding sites.

    Purpose of the Study:

    • To describe the mechanism and application of competitive ELISA (cELISA) for quantifying small-molecule antigens.
    • To highlight the inverse relationship between signal intensity and antigen concentration in cELISA.

    Main Methods:

    • Immobilization of either antibody or antigen onto microtiter plate wells.
    • Competition between labeled analyte (antigen or antibody) and unlabeled analyte in the test sample for binding to the immobilized molecule.
    • Detection of bound labeled analyte using an enzyme-linked secondary antibody or direct labeling, followed by substrate addition and signal measurement.

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    Main Results:

    • The signal generated is inversely proportional to the concentration of the analyte in the test sample.
    • Higher concentrations of antigen in the sample lead to less labeled antigen binding, resulting in a weaker signal.
    • Direct cELISAs utilize labeled antigen or antibody, while indirect configurations employ reporter-labeled secondary antibodies.

    Conclusions:

    • Competitive ELISA is highly effective for determining the concentration of small-molecule antigens within complex sample matrices.
    • The assay provides a sensitive method for analyte quantification based on signal inhibition.
    • Understanding the competitive binding dynamics is crucial for accurate interpretation of cELISA results.