Hybridoma Screening by Antibody Capture: A High-Throughput Western Blot Assay

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Edward A Greenfield

    Cold Spring Harbor Protocols
    |November 2, 2021
    PubMed

    Antibody capture assays are often the easiest and most convenient of the hybridoma screening methods. In this procedure, proteins in solution or in a cell lysate are separated according to size by gel electrophoresis and then transferred by blotting to a nitrocellulose sheet. Antigen bound to the solid substrate is incubated with the primary antibody, and the resultant antibody-antigen complexes are detected by a horseradish peroxidase (HRP)-conjugated secondary antibody and a chemiluminescent substrate for HRP.

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