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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.
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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Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

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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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Related Experiment Video

Updated: Mar 3, 2026

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

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Indirect Immunometric ELISA.

Thomas O Kohl, Carl A Ascoli

    Cold Spring Harbor Protocols
    |May 3, 2017
    PubMed
    Summary
    This summary is machine-generated.

    This assay enables the immunometric determination of reagents like antibodies and analytes. It is a precursor to sandwich enzyme-linked immunosorbent assays (ELISA) for antigen quantification.

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    Area of Science:

    • Biochemistry
    • Immunology
    • Assay Development

    Background:

    • Accurate quantification of analytes and reagents is crucial for immunoassay development.
    • Optimization of antibody concentrations is essential for sensitive and specific antigen measurement.
    • Existing methods may require adaptation for different reporter systems.

    Purpose of the Study:

    • To describe an assay for the immunometric determination of assay-associated reagents.
    • To provide a protocol that can precede the development of sandwich enzyme-linked immunosorbent assays (ELISA).
    • To detail materials and equipment for chromogenic substrate measurement, adaptable for other reporters.

    Main Methods:

    • Immunometric determination of capture antibodies, detection antibodies, and analytes.
    • Protocol for measuring chromogenic substrate development.
    • Adaptable methodology for chemiluminescent- and fluorescent-labeled reporters.

    Main Results:

    • The assay facilitates the determination of key immunoassay components.
    • The protocol supports the optimization of antibody concentrations for quantitative antigen measurement.
    • The method is versatile and can be adapted for various detection systems.

    Conclusions:

    • This assay is a valuable tool for characterizing reagents in immunoassay development.
    • It serves as a foundational step for creating optimized sandwich ELISA protocols.
    • The described methodology offers flexibility for different reporter-based detection strategies.