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Related Concept Videos

The Proteasome01:13

The Proteasome

Eukaryotic cells can degrade proteins through several pathways. One of the most important among these is the ubiquitin-proteasome pathway. It helps the cell eliminate the misfolded, damaged, or unwarranted cytoplasmic proteins in a highly specific manner.
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. This involves participation of a series of enzymes including— E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3 (ubiquitin...
The Proteasome02:18

The Proteasome

Eukaryotic cells can degrade proteins through several pathways. One of the most important amongst these is the ubiquitin-proteasome pathway. It helps the cell eliminate the misfolded, damaged, or unwarranted cytoplasmic proteins in a highly specific manner.
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. A series of enzymes carry out the ubiquitination of the target proteins - E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3...
The Proteasome Structure01:17

The Proteasome Structure

The ubiquitin-proteasome pathway is a well-known mechanism utilized by eukaryotic cells to remove cytoplasmic proteins that are misfolded, damaged, or no longer needed. In this pathway, the protein that needs to be eliminated undergoes a process called ubiquitination, where a chain of ubiquitin molecules is attached to the 48th lysine residue of the target protein. This ubiquitin modification helps the proteasome distinguish between a target protein and a healthy protein.
The proteasome is an...

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Related Experiment Video

Updated: May 23, 2026

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

Unmasking hidden epitopes with proteases.

Ed Harlow, David Lane

    CSH Protocols
    |April 10, 2012
    PubMed
    Summary
    This summary is machine-generated.

    Fixation often hides epitopes, but gentle protease treatment can re-expose them for antibody access. Using characterized proteases like trypsin offers controlled antigen retrieval, though artifacts are possible.

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    Last Updated: May 23, 2026

    A High Throughput MHC II Binding Assay for Quantitative Analysis of Peptide Epitopes
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    Published on: March 25, 2014

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

    • Immunohistochemistry
    • Molecular Biology

    Background:

    • Tissue fixation can obscure antigenic sites (epitopes), hindering antibody binding and detection.
    • Proteolytic digestion offers a method to unmask these hidden epitopes, improving antigen accessibility.

    Purpose of the Study:

    • To describe a method for re-exposing masked epitopes using protease incubation.
    • To highlight the use of characterized proteases for controlled antigen retrieval.

    Main Methods:

    • Incubation of fixed tissues with proteases (e.g., pronase, trypsin).
    • Enzymatic removal of obstructing structures to allow antibody penetration.

    Main Results:

    • Protease treatment successfully re-exposed previously inaccessible epitopes.
    • Trypsin provided a more controlled enzymatic reaction compared to crude protease preparations.

    Conclusions:

    • Gentle protease incubation is an effective antigen retrieval technique for immunohistochemistry.
    • Careful selection of proteases and awareness of potential artifacts are crucial for reliable results.