Related Experiment Video

Updated: Sep 20, 2025

Single-cell Screening Method for the Selection and Recovery of Antibodies with Desired Specificities from Enriched Human Memory B Cell Populations
09:07

Single-cell Screening Method for the Selection and Recovery of Antibodies with Desired Specificities from Enriched Human Memory B Cell Populations

Published on: August 22, 2019

11.0K

Hybridoma Screening by Antibody Capture: Immunohistochemistry

Edward A Greenfield

    Cold Spring Harbor Protocols
    |June 7, 2022
    PubMed

    Insights

    Immunohistochemistry uses antibodies to locate antigens in tissues. Proper tissue preparation, including paraffin removal and rehydration, is crucial for antibody access and accurate staining results.

    Area of Science:

    • Cellular Biology
    • Immunology
    • Histology

    Background:

    • Immunohistochemistry (IHC) is a key technique for visualizing antigen localization within tissues.
    • Hybridoma technology produces monoclonal antibodies essential for specific antigen detection in IHC.

    Purpose of the Study:

    • To outline the critical steps for preparing fixed and embedded tissue sections for immunohistochemical staining.
    • To ensure optimal antibody penetration and antigen accessibility in histological samples.

    Main Methods:

    • Tissue preparation involves deparaffinization, rehydration, and enzymatic or chemical digestion.
    • Screening of hybridoma supernatants using immunohistochemistry on prepared tissue sections.

    Main Results:

    • Successful antigen localization depends on effective removal of embedding media and proper tissue rehydration.
    • Optimized tissue digestion enhances antibody access to target antigens, improving staining sensitivity and specificity.

    Conclusions:

    • Standardized tissue processing protocols are essential for reliable immunohistochemistry.
    • Effective antigen retrieval methods are critical for successful subcellular localization studies using IHC.

    More Related Videos

    Author Spotlight: Efficient Detection of Immune Cell-Infiltration in Cancer Tissues Using Fluorescent Immunohistochemistry04:21

    Author Spotlight: Efficient Detection of Immune Cell-Infiltration in Cancer Tissues Using Fluorescent Immunohistochemistry

    Published on: January 26, 2024

    920
    Generation of Monoclonal Antibodies Against Natural Products12:15

    Generation of Monoclonal Antibodies Against Natural Products

    Published on: April 6, 2019

    11.1K

    Related Concept Videos

    Hybridoma Technology01:31

    Hybridoma Technology

    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,...
    15.4K
    Immunocytochemistry and Immunohistochemistry01:22

    Immunocytochemistry and Immunohistochemistry

    Immunocytochemistry (ICC) and immunohistochemistry (IHC) are techniques that use antibodies to check for specific proteins or antigens in a sample. The technique was first published by Albert Coons in 1941 to detect the presence of pneumococcal antigen in tissue sections from mice infected with Pneumococcus. Immunocytochemistry helps localization of proteins or antigens in individual cells like blood cells, stem cells, etc., while immunohistochemistry does the same for tissue samples.
    These...
    12.0K