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Related Experiment Video

Updated: May 23, 2026

Detection of Axonally Localized mRNAs in Brain Sections Using High-Resolution In Situ Hybridization
11:24

Detection of Axonally Localized mRNAs in Brain Sections Using High-Resolution In Situ Hybridization

Published on: June 17, 2015

Unmasking hidden epitopes using the microwave oven.

Ed Harlow, David Lane

    CSH Protocols
    |April 10, 2012
    PubMed
    Summary

    Antigen retrieval methods, like microwave heating, unmask antigens for antibody detection but may cause false positives. These techniques are antibody-specific and not suitable for quantitative analysis.

    Area of Science:

    • Immunohistochemistry
    • Biochemical techniques

    Background:

    • Antigen masking by fixation hinders antibody binding in immunohistochemistry.
    • Several methods exist to reverse fixation-induced antigen masking.

    Purpose of the Study:

    • To describe a microwave oven-based method for antigen retrieval.
    • To highlight potential artifacts and limitations of antigen retrieval techniques.

    Main Methods:

    • Utilizing microwave heating for extended periods to disrupt blocking subcellular structures.
    • Applying antigen retrieval methods specific to antibody/antigen combinations.

    Main Results:

    • Microwave heating can break down structures that block antibody access.
    • Antigen retrieval methods can introduce artifactual false-positive staining.

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    A High Throughput MHC II Binding Assay for Quantitative Analysis of Peptide Epitopes
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    A High Throughput MHC II Binding Assay for Quantitative Analysis of Peptide Epitopes

    Published on: March 25, 2014

    Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope
    08:09

    Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope

    Published on: March 24, 2017

    Related Experiment Videos

    Last Updated: May 23, 2026

    Detection of Axonally Localized mRNAs in Brain Sections Using High-Resolution In Situ Hybridization
    11:24

    Detection of Axonally Localized mRNAs in Brain Sections Using High-Resolution In Situ Hybridization

    Published on: June 17, 2015

    A High Throughput MHC II Binding Assay for Quantitative Analysis of Peptide Epitopes
    07:59

    A High Throughput MHC II Binding Assay for Quantitative Analysis of Peptide Epitopes

    Published on: March 25, 2014

    Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope
    08:09

    Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope

    Published on: March 24, 2017

    Conclusions:

    • Antigen retrieval methods should be used cautiously and avoided if possible.
    • These methods are not suitable for generating quantitative data due to potential artifacts.