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Related Concept Videos

Autophagy01:27

Autophagy

Autophagy is a self-digesting process by which a cell protects itself from threats both within and outside the cell, ranging from abnormal proteins to invading bacteria. In this process, obsolete components of the cell and invading microbes are degraded by hydrolytic enzymes active in an acidic environment of the lysosomal lumen.
An autophagic pathway consists of a series of signaling events activated in response to diverse stress and physiological conditions such as food deprivation,...

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Related Experiment Video

Updated: Jun 15, 2026

Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry
11:39

Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry

Published on: July 21, 2017

Autophagy: assays and artifacts.

Sandra Barth1, Danielle Glick, Kay F Macleod

  • 1Ben May Department for Cancer Research, Gordon Center for Integrative Sciences, University of Chicago, IL, USA.

The Journal of Pathology
|March 13, 2010
PubMed
Summary

Autophagy monitoring assays are crucial for understanding its role in health and disease. This review critically examines current methods for measuring autophagy in cells and animals, highlighting limitations and the need for improved techniques.

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The Lactate Dehydrogenase Sequestration Assay — A Simple and Reliable Method to Determine Bulk Autophagic Sequestration Activity in Mammalian Cells
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The Lactate Dehydrogenase Sequestration Assay — A Simple and Reliable Method to Determine Bulk Autophagic Sequestration Activity in Mammalian Cells

Published on: July 27, 2018

Related Experiment Videos

Last Updated: Jun 15, 2026

Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry
11:39

Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry

Published on: July 21, 2017

The Lactate Dehydrogenase Sequestration Assay — A Simple and Reliable Method to Determine Bulk Autophagic Sequestration Activity in Mammalian Cells
09:34

The Lactate Dehydrogenase Sequestration Assay — A Simple and Reliable Method to Determine Bulk Autophagic Sequestration Activity in Mammalian Cells

Published on: July 27, 2018

Area of Science:

  • Cellular Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Autophagy is a conserved cellular degradation process essential for development and disease.
  • Current methods like LC3B-II detection and electron microscopy have limitations.
  • Measuring autophagic flux, not just static levels, is increasingly important.

Purpose of the Study:

  • To provide a critical overview of existing autophagy detection methodologies.
  • To address the need for reliable and quantitative assays for autophagy research.
  • To discuss challenges and advancements in measuring autophagy in vitro and in vivo.

Main Methods:

  • Review of current literature on autophagy detection assays.
  • Critical analysis of western blot, fluorescence studies, and electron microscopy.
  • Discussion of novel and modified approaches for measuring autophagic flux.

Main Results:

  • Traditional methods like LC3B-II detection have limitations due to variable expression and potential interference from tagged proteins.
  • Electron microscopy is useful for visualizing autophagosomes but not for quantitative flux analysis.
  • Newer methods are being developed to better assess autophagic flux.

Conclusions:

  • Existing autophagy assays require careful validation and interpretation.
  • There is a significant need for improved, quantitative methods to monitor autophagy and autophagic flux.
  • Accurate measurement of autophagy is critical for advancing research in development and disease.