Journal
/
Biochemistry
/
Analyzing the Interaction of Fluorescent-Labeled Proteins with Artificial Phospholipid Microvesicles using Quantitative Flow Cytometry
JoVE Journal
Biochemistry
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biochemistry
Analyzing the Interaction of Fluorescent-Labeled Proteins with Artificial Phospholipid Microvesicles using Quantitative Flow Cytometry

Analyzing the Interaction of Fluorescent-Labeled Proteins with Artificial Phospholipid Microvesicles using Quantitative Flow Cytometry

DOI:
10.3791/63459-v

•

08:26 min

•

April 06, 2022

•

, , , ,
1Center for Theoretical Problems of Physicochemical Pharmacology,Russian Academy of Sciences, 2National Medical Research Center of Pediatric Hematology, Oncology and Immunology named after Dmitry Rogachev, 3Faculty of Physics,Lomonosov Moscow State University, 4Faculty of Biological and Medical Physics,Moscow Institute of Physics and Technology

Chapters

  • 00:04Introduction
  • 00:58Kinetic Binding Experiments
  • 02:13Equilibrium Binding Experiments
  • 02:45Analysis of Flow Cytometry Data
  • 05:13Converting Fluorescence Intensity to the Mean Number of Binding Sites
  • 07:05Results: Specific Binding of Fluorescent-Labeled Coagulation Factor X or fX to Artificial Phospholipid Vesicles
  • 07:42Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we describe a set of methods for characterizing the interaction of proteins with membranes of cells or microvesicles.

Tags

Keywords: Flow CytometryPhospholipid MicrovesiclesFluorescent-labeled ProteinsKinetic BindingEquilibrium BindingCoagulation Factor XTyrode’s BufferFX FDMembrane InteractionsQuantitative Analysis

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/63459?article_type=v

Mimicking the Function of Signaling Proteins: Toward Artificial Signal Transduction Therapy
check_url/63459?article_type=v

A Fluorescence-based Assay of Phospholipid Scramblase Activity
check_url/63459?article_type=v

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
check_url/63459?article_type=v

Dissipative Microgravimetry to Study the Binding Dynamics of the Phospholipid Binding Protein Annexin A2 to Solid-supported Lipid Bilayers Using a Quartz Resonator
check_url/63459?article_type=v

Localization of SUMO-modified Proteins Using Fluorescent Sumo-trapping Proteins
check_url/63459?article_type=v

Quantitative Analysis of the Cellular Lipidome of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry
check_url/63459?article_type=v

Transverse Sectioning of Mature Rice (Oryza sativa L.) Kernels for Scanning Electron Microscopy Imaging Using Pipette Tips as Immobilization Support
check_url/63459?article_type=v

Quantitative Metabolomics of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry
check_url/63459?article_type=v

A Robust Single-Particle Cryo-Electron Microscopy (cryo-EM) Processing Workflow with cryoSPARC, RELION, and Scipion
check_url/63459?article_type=v

Rapid Assessment of Membrane Protein Quality by Fluorescent Size Exclusion Chromatography

Read Article