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Related Concept Videos

Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

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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.
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A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
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Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.
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Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
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Immunoassays.

Thomas O Kohl, Carl A Ascoli

    Cold Spring Harbor Protocols
    |July 7, 2017
    PubMed
    Summary
    This summary is machine-generated.

    Enzyme immunoassays (EIA) are powerful immunochemical techniques for detecting and quantifying antigens and antibodies. This work details the development and optimization of enzyme-linked immunosorbent assays (ELISA) for sensitive biological analysis.

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    Area of Science:

    • Immunochemistry
    • Biotechnology
    • Analytical Chemistry

    Background:

    • Enzyme immunoassay (EIA) is a robust immunochemical technique developed in the 1970s.
    • EIAs are routinely employed in laboratory diagnostics and analyses for detecting and quantifying analytes.
    • These assays offer high sensitivity for measuring low concentrations of antigens and antibodies.

    Purpose of the Study:

    • To detail the development and optimization of enzyme-linked immunosorbent assays (ELISA).
    • To highlight ELISA as a versatile tool for antigen and antibody quantification in biological and biotechnological applications.

    Main Methods:

    • Development and optimization of enzyme-linked immunosorbent assays (ELISA).
    • Utilizing enzyme-, chemiluminescence-, or fluorescence-based reporters in plate-based immunoassays.

    Main Results:

    • Demonstration of ELISA's capability for rapid quantification of analytes at very low concentrations.
    • Highlighting the sensitivity and valuable information provided by EIAs.

    Conclusions:

    • Enzyme-linked immunosorbent assays (ELISA) are highly sensitive and versatile tools.
    • ELISA is amenable to standardization, automation, and large-scale sampling for biological and diagnostic applications.