Related Experiment Video
Updated: May 1, 2026
06:59
The Nijmegen Hemostasis Assay: Simultaneous Fluorogenic Measurement of Thrombin and Plasmin Generation in a Single Well
Published on: February 27, 2026
174
Whole blood thrombin generation in Bmal1-deficient mice
M Ninivaggi, H Kelchtermans, M J Kuijpers
1B. de Laat, PhD, Synapse bv, Oxfordlaan 70, 6229 EV Maastricht, The Netherlands, Tel.: +31 43 3885895, Fax: +31 43 3884159,
Thrombosis and Haemostasis
|April 18, 2014
Summary
A new whole blood assay accurately measures thrombin generation (TG) in mice. This assay revealed a prothrombotic phenotype in premature aging Bmal1-knockout mice, indicating increased clotting potential.
Area of Science:
- Hemostasis and Thrombosis
- Aging Research
- Biomedical Assay Development
Background:
- The Calibrated Automated Thrombogram (CAT) assay is a sensitive tool for assessing hemostatic system function.
- Existing methods require plasma, necessitating a simpler whole blood approach.
- Mouse models of aging are crucial for understanding age-related hemostatic changes.
Purpose of the Study:
- To validate a novel whole blood thrombin generation (TG) assay in mouse blood.
- To investigate TG dynamics in a mouse model exhibiting premature aging symptoms.
- To correlate TG measurements with fibrin network morphology.
Main Methods:
- Developed and validated a whole blood TG assay using small blood aliquots.
- Assayed TG in brain and muscle ARNT-like protein-1 (Bmal1)-deficient (knockout) and wild-type mice.
- Initiated TG with calcium, tissue factor, and a thrombin substrate.
- Analyzed fibrin structure using scanning electron microscopy (SEM).
Main Results:
- The whole blood TG assay demonstrated high reproducibility with intra-assay variations ≤10%.
- Bmal1-knockout mice exhibited significantly higher endogenous thrombin potential (ETP) and peak TG compared to wild-type controls.
- SEM revealed a denser fibrin network with smaller pores in Bmal1-knockout mice.
Conclusions:
- The developed whole blood TG assay is a reproducible and sensitive method for mouse blood.
- This assay successfully identified a prothrombotic phenotype in Bmal1-knockout mice.
- The findings suggest a link between Bmal1 deficiency, accelerated aging, and altered hemostasis.
