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Related Concept Videos

Hybridoma Technology01:31

Hybridoma Technology

15.6K
Hybridoma technology is used for the large-scale production of monoclonal antibodies. Monoclonal antibodies bind to only a single antigenic determinant or epitope. Such antibodies are used in research, diagnostics, and disease therapy. The hybridoma technology established in 1975 by Georges Köhler and Cesar Milstein was awarded the Nobel Prize in Medicine in 1984 for revolutionizing research and therapy.
Hybridoma Selection
Commonly used fusion techniques — electroporation,...
15.6K
Western Blotting01:15

Western Blotting

18.0K
Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.
18.0K

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Related Experiment Video

Updated: Oct 8, 2025

Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope
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Hybridoma Screening by Antigen Capture: Reverse Dot Blot.

Edward A Greenfield

    Cold Spring Harbor Protocols
    |January 5, 2022
    PubMed
    Summary

    A dot blot assay effectively screens for productive hybridoma clones by capturing monoclonal antibodies. This method uses nitrocellulose membranes and horseradish peroxidase detection for hybridoma screening.

    Area of Science:

    • Immunology
    • Biotechnology

    Background:

    • Hybridoma technology is crucial for monoclonal antibody production.
    • Screening for productive hybridoma clones is essential for efficient antibody generation.

    Purpose of the Study:

    • To describe the dot blot assay for assessing hybridoma productivity.
    • To provide a method for screening hybridoma fusion and subclone plates.

    Main Methods:

    • Nitrocellulose membranes are coated with anti-mouse immunoglobulin.
    • Hybridoma tissue culture supernatant is applied to the membrane.
    • Antibodies are detected using horseradish peroxidase (HRP) conjugated secondary antibodies.

    Main Results:

    • The dot blot assay allows for the visualization of captured monoclonal antibodies.

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    Generation of Monoclonal Antibodies Against Natural Products
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  • This method enables the identification of high-producing hybridoma clones.

    • Conclusions:

    • The dot blot assay is a reliable and widely used method for determining hybridoma productivity.
    • It serves as an efficient screening tool for selecting productive hybridoma clones.