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Cells 2017, 6(3), 17;

Standard Immunohistochemical Assays to Assess Autophagy in Mammalian Tissue

Laboratory of Physiopharmacology, University of Antwerp, Universiteitsplein 1, B-2610 Antwerp, Belgium
Author to whom correspondence should be addressed.
Received: 24 May 2017 / Revised: 20 June 2017 / Accepted: 26 June 2017 / Published: 30 June 2017
(This article belongs to the Special Issue Assays to Monitor Autophagy in Model Systems)
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Autophagy is a highly conserved lysosomal degradation pathway with major impact on diverse human pathologies. Despite the development of different methodologies to detect autophagy both in vitro and in vivo, monitoring autophagy in tissue via immunohistochemical techniques is hampered due to the lack of biomarkers. Immunohistochemical detection of a punctate pattern of ATG8/MAP1LC3 proteins is currently the most frequently used approach to detect autophagy in situ, but it depends on a highly sensitive detection method and is prone to misinterpretation. Moreover, reliable MAP1LC3 immunohistochemical staining requires correct tissue processing and high-quality, isoform-specific antibodies. Immunohistochemical analysis of other autophagy-related protein targets such as SQSTM1, ubiquitin, ATG5 or lysosomal proteins is not recommended as marker for autophagic activity in tissue for multiple reasons including aspecific labeling of cellular structures and a lack of differential protein expression during autophagy initiation. To better understand the role of autophagy in human disease, novel biomarkers for visualization of the autophagic process with standard histology techniques are urgently needed. View Full-Text
Keywords: autophagy; immunohistochemistry; MAP1LC3; SQSTM1; autophagosome autophagy; immunohistochemistry; MAP1LC3; SQSTM1; autophagosome

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Martinet, W.; Roth, L.; De Meyer, G.R.Y. Standard Immunohistochemical Assays to Assess Autophagy in Mammalian Tissue. Cells 2017, 6, 17.

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