Analysis of macroautophagy by immunohistochemistry
(Macro)Autophagy is a phylogenetically conserved membrane-trafficking process that functions to deliver cytoplasmic cargoes to lysosomes for digestion. The process is a major mechanism for turnover of cellular constituents and is therefore critical for maintaining cellular homeostasis. Macroautophag...
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| Format: | Online |
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Landes Bioscience
2012
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| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3427261/ |
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pubmed-3427261 |
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pubmed-34272612012-08-27 Analysis of macroautophagy by immunohistochemistry Rosenfeldt, Mathias T. Nixon, Colin Liu, Emma Mah, Li Yen Ryan, Kevin M. Protocol (Macro)Autophagy is a phylogenetically conserved membrane-trafficking process that functions to deliver cytoplasmic cargoes to lysosomes for digestion. The process is a major mechanism for turnover of cellular constituents and is therefore critical for maintaining cellular homeostasis. Macroautophagy is characteristically distinct from other forms of autophagy due to the formation of double-membraned vesicles termed autophagosomes which encapsulate cargoes prior to fusion with lysosomes. Autophagosomes contain an integral membrane-bound form (LC3-II) of the microtubule-associated protein 1 light chain 3 β (MAP1LC3B), which has become a gold-standard marker to detect accumulation of autophagosomes and thereby changes in macroautophagy. Due to the role played by macroautophagy in various diseases, the detection of autophagosomes in tissue sections is frequently desired. To date, however, the detection of endogenous LC3-II on paraffin-embedded tissue sections has proved problematic. We report here a simple, optimized and validated method for the detection of LC3-II by immunohistochemistry in human and mouse tissue samples that we believe will be a useful resource for those wishing to study macroautophagy ex vivo. Landes Bioscience 2012-06-01 /pmc/articles/PMC3427261/ /pubmed/22562096 http://dx.doi.org/10.4161/auto.20186 Text en Copyright © 2012 Landes Bioscience http://creativecommons.org/licenses/by-nc/3.0/ This is an open-access article licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License. The article may be redistributed, reproduced, and reused for non-commercial purposes, provided the original source is properly cited. |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
Rosenfeldt, Mathias T. Nixon, Colin Liu, Emma Mah, Li Yen Ryan, Kevin M. |
| spellingShingle |
Rosenfeldt, Mathias T. Nixon, Colin Liu, Emma Mah, Li Yen Ryan, Kevin M. Analysis of macroautophagy by immunohistochemistry |
| author_facet |
Rosenfeldt, Mathias T. Nixon, Colin Liu, Emma Mah, Li Yen Ryan, Kevin M. |
| author_sort |
Rosenfeldt, Mathias T. |
| title |
Analysis of macroautophagy by immunohistochemistry |
| title_short |
Analysis of macroautophagy by immunohistochemistry |
| title_full |
Analysis of macroautophagy by immunohistochemistry |
| title_fullStr |
Analysis of macroautophagy by immunohistochemistry |
| title_full_unstemmed |
Analysis of macroautophagy by immunohistochemistry |
| title_sort |
analysis of macroautophagy by immunohistochemistry |
| description |
(Macro)Autophagy is a phylogenetically conserved membrane-trafficking process that functions to deliver cytoplasmic cargoes to lysosomes for digestion. The process is a major mechanism for turnover of cellular constituents and is therefore critical for maintaining cellular homeostasis. Macroautophagy is characteristically distinct from other forms of autophagy due to the formation of double-membraned vesicles termed autophagosomes which encapsulate cargoes prior to fusion with lysosomes. Autophagosomes contain an integral membrane-bound form (LC3-II) of the microtubule-associated protein 1 light chain 3 β (MAP1LC3B), which has become a gold-standard marker to detect accumulation of autophagosomes and thereby changes in macroautophagy. Due to the role played by macroautophagy in various diseases, the detection of autophagosomes in tissue sections is frequently desired. To date, however, the detection of endogenous LC3-II on paraffin-embedded tissue sections has proved problematic. We report here a simple, optimized and validated method for the detection of LC3-II by immunohistochemistry in human and mouse tissue samples that we believe will be a useful resource for those wishing to study macroautophagy ex vivo. |
| publisher |
Landes Bioscience |
| publishDate |
2012 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3427261/ |
| _version_ |
1611552121535070208 |