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Related Concept Videos

Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

18.0K
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...
18.0K

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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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Direct and Indirect Cell-Based Enzyme-Linked Immunosorbent Assay.

Thomas O Kohl, Carl A Ascoli

    Cold Spring Harbor Protocols
    |May 3, 2017
    PubMed
    Summary

    Cell-based enzyme-linked immunosorbent assays (ELISA) offer rapid detection of cell surface antigens. This immunoassay method aids in identifying antibodies for research and diagnostics.

    Area of Science:

    • Immunology
    • Cell Biology
    • Biotechnology

    Background:

    • Cell-based enzyme-linked immunosorbent assays (ELISA) are increasingly utilized for detecting cell surface antigens and receptors.
    • Traditional methods often require complex sample preparation or specialized equipment.

    Purpose of the Study:

    • To describe the methodology and applications of cell-based ELISA for antibody screening and characterization.
    • To highlight its utility as a surrogate for immunohistochemistry (IHC) antibody validation.

    Main Methods:

    • Cells are adhered to microtiter plates, fixed, and incubated with either hybridoma supernatants or labeled antibodies.
    • Washing steps remove unbound antibodies, followed by addition of a secondary antibody (indirect approach) and substrate.
    • Substrate hydrolysis is measured spectrophotometrically to quantify antigen expression.

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    Electrowetting-based Digital Microfluidics Platform for Automated Enzyme-linked Immunosorbent Assay
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    Main Results:

    • Cell-based ELISA provides sensitive detection of cell surface antigen expression, comparable to flow cytometry.
    • The indirect approach effectively screens hybridoma supernatants for specific antibody reactivity.
    • Multiplex formats are achievable using different reporters for simultaneous analysis.

    Conclusions:

    • Cell-based ELISA is a versatile and sensitive immunoassay for evaluating cell surface antigen expression.
    • It serves as a valuable tool for antibody validation, hybridoma screening, and diagnostic assay development.
    • Its adaptability supports diverse research applications in immunology and cell biology.