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

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

2.2K
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
2.2K

You might also read

Related Articles

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

Sort by
Same author

A primordial synaptotagmin-independent function of complexin in regulated exocytosis.

Nature communications·2026
Same author

Evolution from Monolayers to Two-Dimensional Heterostructures for Enhanced Hydrogen Evolution Reaction: A Theoretical Study.

Molecules (Basel, Switzerland)·2026
Same author

Seasonal variation of fish biodiversity in Xisha Islands based on dual-marker eDNA metabarcoding.

Marine environmental research·2026
Same author

Data-Independent Acquisition-Based Quantitative Proteomics, Nontargeted Metabolomics, and Protein-Ligand Interaction Reveal the Mechanism of Baicalin Inhibiting β<i>-</i>hemolytic/cytolytic Activities of <i>Streptococcus agalactiae</i>.

Journal of agricultural and food chemistry·2026
Same author

Regulatory Effects of a Lipid-Lowering Strain <i>Lactobacillus plantarum</i> 58 Isolated From Dregs Vinegar on Metabolism-Related Gene Expression, Gut Microbiota, and Metabolic Biomarkers of Hybrid Grouper Under High-Fat Diets.

Aquaculture nutrition·2026
Same author

Two-dimensional SnSe<sub>2</sub>/Bi<sub>2</sub>Se<sub>3</sub> vertically stacked heterostructures for enhanced photoresponse through tuning layer coverage.

iScience·2026

Related Experiment Video

Updated: Sep 5, 2025

Imaging FITC-dextran as a Reporter for Regulated Exocytosis
04:50

Imaging FITC-dextran as a Reporter for Regulated Exocytosis

Published on: June 20, 2018

12.8K

Endocytosis Assays Using Cleavable Fluorescent Dyes.

Shifeng Wang1,2, Chun Wan3, Galen T Squiers3

  • 1Department of Chinese Medicine Information Science, Beijing University of Chinese Medicine, Beijing, China. shifeng.wang@colorado.edu.

Methods in Molecular Biology (Clifton, N.J.)
|July 12, 2022
PubMed
Summary
This summary is machine-generated.

This chapter details methods to quantify endocytosis of soluble and transmembrane proteins using fluorescent dyes and antibodies. It also presents strategies to investigate the role of clathrin-mediated endocytosis (CME) in cargo internalization.

Keywords:
AAGABCargo proteinClathrin-mediated endocytosisEndocytosisGLUT4Membrane traffickingTransferrinTransferrin receptor

More Related Videos

Extracellular Vesicle Uptake Assay via Confocal Microscope Imaging Analysis
08:32

Extracellular Vesicle Uptake Assay via Confocal Microscope Imaging Analysis

Published on: February 14, 2022

8.1K
A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts
08:43

A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts

Published on: December 1, 2018

11.5K

Related Experiment Videos

Last Updated: Sep 5, 2025

Imaging FITC-dextran as a Reporter for Regulated Exocytosis
04:50

Imaging FITC-dextran as a Reporter for Regulated Exocytosis

Published on: June 20, 2018

12.8K
Extracellular Vesicle Uptake Assay via Confocal Microscope Imaging Analysis
08:32

Extracellular Vesicle Uptake Assay via Confocal Microscope Imaging Analysis

Published on: February 14, 2022

8.1K
A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts
08:43

A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts

Published on: December 1, 2018

11.5K

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Endocytosis is a fundamental cellular process responsible for internalizing surface and extracellular materials.
  • Understanding endocytic mechanisms is crucial for various biological processes, including nutrient uptake, signal transduction, and pathogen entry.

Purpose of the Study:

  • To describe quantitative assays for measuring the endocytosis of soluble and transmembrane cargo proteins.
  • To present strategies for investigating the role of clathrin-mediated endocytosis (CME) in cargo internalization.

Main Methods:

  • Utilizing cleavable fluorescent dyes to label cargo proteins for quantitative measurement.
  • Employing antibodies to recognize and track transmembrane cargo proteins.
  • Applying confocal imaging and flow cytometry to quantify internalized cargoes after removing surface-bound labels.
  • Using small molecule inhibitors of CME and AAGAB gene knockout (KO) to assess CME's role.

Main Results:

  • Established quantitative assays for measuring endocytosis of diverse cargo types.
  • Demonstrated methodologies to differentiate between surface-bound and internalized cargo.
  • Provided tools to investigate the specific contribution of clathrin-mediated endocytosis.

Conclusions:

  • The described assays provide robust methods for quantifying endocytosis.
  • These methods facilitate the study of cargo internalization pathways, including the role of CME.
  • Investigating CME using genetic and chemical tools offers insights into cellular trafficking.