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Related Experiment Video

Updated: Jul 5, 2025

Author Spotlight: High-Sensitivity Tissue Factor Activity Assay for Plasma Diagnosis
03:53

Author Spotlight: High-Sensitivity Tissue Factor Activity Assay for Plasma Diagnosis

Published on: December 29, 2023

811

Extracellular Vesicle Tissue Factor Activity Assay.

Yohei Hisada1, Nigel Mackman2

  • 1Division of Hematology, UNC Blood Research Center, University of North Carolina at Chapel Hill.

Journal of Visualized Experiments : Jove
|January 15, 2024
PubMed
Summary
This summary is machine-generated.

This study presents a new assay to measure tissue factor (TF)-positive extracellular vesicles (EVs) in plasma. The developed activity-based assay offers higher sensitivity and specificity for detecting TF-positive EVs, crucial in various disease states.

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Related Experiment Videos

Last Updated: Jul 5, 2025

Author Spotlight: High-Sensitivity Tissue Factor Activity Assay for Plasma Diagnosis
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Author Spotlight: High-Sensitivity Tissue Factor Activity Assay for Plasma Diagnosis

Published on: December 29, 2023

811
Flow Cytometry Analysis of Tissue Factor Expression in Human Platelets
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Author Spotlight: Deciphering Coagulation Disorders in Traumatic Brain Injury Patients
04:56

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Published on: August 4, 2023

775

Area of Science:

  • Biochemistry
  • Hematology
  • Cell Biology

Background:

  • Tissue factor (TF) is central to initiating the coagulation cascade.
  • TF is released on extracellular vesicles (EVs) into circulation.
  • Elevated TF-positive (TF+) EVs correlate with diseases, thrombosis, and mortality.

Purpose of the Study:

  • To develop and validate a sensitive and specific assay for measuring TF activity on EVs.
  • To improve upon existing antigen-based assays for TF+ EV detection.

Main Methods:

  • Developed a two-stage FXa generation assay to measure TF activity on EVs.
  • Utilized FVIIa, FX, and calcium to quantify TF-dependent FXa generation.
  • Employed a chromogenic substrate and a standard curve for TF quantification.

Main Results:

  • The in-house assay accurately measures TF-dependent FXa generation.
  • Demonstrated higher sensitivity and specificity compared to a commercial TF activity assay.
  • Distinguished TF-dependent from TF-independent FXa generation using an anti-TF antibody.

Conclusions:

  • The novel in-house assay provides a more sensitive and specific method for quantifying TF activity on EVs.
  • This assay can improve the assessment of TF+ EVs in disease diagnosis and monitoring.
  • Activity-based assays are superior to antigen-based assays for TF+ EV measurement.