Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Autophagy01:27

Autophagy

6.5K
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,...
6.5K
Delivery Pathways to the Lysosome01:36

Delivery Pathways to the Lysosome

10.6K
Eukaryotic cells use different mechanisms to eliminate toxic waste obsolete and worn-out substances. Lysosomes play a pivotal role in this, and hence, these substances are carried to the lysosome from other parts of the cell and extracellular space through different pathways. The most elaborately studied pathways to the lysosome are the endocytic pathways.
Endocytosis
In endocytosis, the cell membrane takes up macromolecules and particles from the surrounding medium. Clathrin-mediated...
10.6K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Genomic Prostate Score for Identifying Biologically Low-risk Disease in Patients with Prostate Cancer Undergoing Active Surveillance, Surgery, or Radiotherapy.

European urology oncology·2026
Same author

3D pathology-guided microdissection.

Nature methods·2026
Same author

Assessing the Effects of a 3D Pathology Tissue-Processing Workflow on Downstream Molecular Analyses.

Laboratory investigation; a journal of technical methods and pathology·2026
Same author

Germline polygenic score for prostate cancer aggressiveness.

medRxiv : the preprint server for health sciences·2026
Same author

Identification of small-molecule autophagy activators via GFP-LC3 high-throughput screening and a cargo-based autophagy flux and processing assay.

European journal of pharmacology·2026
Same author

Spatial Robustness of Prostate Cancer Biomarkers Evaluated by Spatial Transcriptomics.

The Prostate·2026

Related Experiment Video

Updated: Apr 18, 2026

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

18.8K

Macroautophagic cargo sequestration assays.

Per O Seglen1, Morten Luhr1, Ian G Mills2

  • 1Centre for Molecular Medicine Norway, Nordic EMBL Partnership, University of Oslo and Oslo University Hospital, N-0318 Oslo, Norway.

Methods (San Diego, Calif.)
|January 11, 2015
PubMed
Summary
This summary is machine-generated.

Monitoring macroautophagy requires functional cargo sequestration assays, as LC3 protein markers are insufficient. The validated LDH sequestration assay measures cargo uptake into autophagic vacuoles, offering a reliable method for studying autophagy.

Keywords:
AutophagyCargo sequestrationLC3LDHLactate dehydrogenaseMacroautophagy

More Related Videos

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

32.6K
A Fluorescence Microscopy Assay for Monitoring Mitophagy in the Yeast Saccharomyces cerevisiae
10:27

A Fluorescence Microscopy Assay for Monitoring Mitophagy in the Yeast Saccharomyces cerevisiae

Published on: July 18, 2011

26.3K

Related Experiment Videos

Last Updated: Apr 18, 2026

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

18.8K
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

32.6K
A Fluorescence Microscopy Assay for Monitoring Mitophagy in the Yeast Saccharomyces cerevisiae
10:27

A Fluorescence Microscopy Assay for Monitoring Mitophagy in the Yeast Saccharomyces cerevisiae

Published on: July 18, 2011

26.3K

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Macroautophagy is a cellular degradation process involving cytoplasmic sequestration and lysosomal breakdown.
  • The autophagy-related protein LC3 is a common marker, but its association with membranes does not guarantee functional cargo sequestration.
  • Existing LC3-based assays lack information on the actual sequestration activity of macroautophagy.

Purpose of the Study:

  • To provide an overview of methods for measuring macroautophagic sequestration activity.
  • To emphasize and validate the Lactate Dehydrogenase (LDH) sequestration assay as a measure of functional macroautophagy.
  • To demonstrate the applicability of the LDH assay in various mammalian cell lines.

Main Methods:

  • Review of historical and current approaches for assessing macroautophagic sequestration.
  • Detailed description and validation of the LDH sequestration assay.
  • Application of the simplified LDH assay to commonly used cell lines for autophagy research.

Main Results:

  • The LDH sequestration assay directly measures the transfer of cytoplasmic cargo (LDH) into autophagic vacuoles.
  • The assay provides a functional readout of macroautophagic activity, independent of LC3.
  • The validated and simplified LDH assay is effective across multiple mammalian cell types.

Conclusions:

  • Traditional LC3-based assays do not fully reflect macroautophagic flux.
  • The LDH sequestration assay offers a robust and validated method to measure functional macroautophagy.
  • This assay is a valuable tool for studying autophagy in various cellular contexts.