Immunohistochemical Detection of the Autophagy Markers LC3 and p62/SQSTM1 in Formalin-Fixed and Paraffin-Embedded Tissue

Methods Mol Biol. 2023:2566:133-139. doi: 10.1007/978-1-0716-2675-7_10.


Autophagy is a highly conserved cellular mechanism of "self-digestion," ensuring cellular homeostasis and playing a role in many diseases including cancer. As a stress response mechanism, it may also be involved in cellular response to therapy. LC3 and Sequestosome 1 (p62/SQSTM1) are among the most widely used markers to monitor autophagy and can be visualized in formalin-fixed and paraffin-embedded tissue by immunohistochemistry. Here we describe a validated staining protocol using an automated staining system available in many routine pathology laboratories, enabling high-throughput staining under standardized conditions.

Keywords: Autophagy; Immunohistochemistry; LC3; Sequestosome 1; p62/SQSTM1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autophagy*
  • Biomarkers
  • Formaldehyde*
  • Immunohistochemistry
  • Microtubule-Associated Proteins / metabolism
  • Paraffin Embedding
  • Sequestosome-1 Protein / metabolism


  • Biomarkers
  • Microtubule-Associated Proteins
  • Sequestosome-1 Protein
  • Formaldehyde