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Updated: Jul 13, 2026
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Use of a Caspase Multiplexing Assay to Determine Apoptosis in a Hypothalamic Cell Model
Published on: April 16, 2014
Two optimized combination assays to examine apoptosis pathways in clinical samples
Mark Hollier1, Toni Whistler, Carolyn Dawson
1Chronic Viral Diseases Branch, Division of Viral and Rickettsial Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30329, USA.
Summary
This study developed a combined assay to measure key apoptosis stages in clinical blood samples. The optimized method efficiently analyzes apoptotic cells and affected blood cell subsets from limited samples.
Area of Science:
- Biochemistry
- Cell Biology
- Immunology
Background:
- Apoptosis (programmed cell death) is frequently altered in various diseases.
- Existing assays for apoptosis are not comprehensive, necessitating multiple tests.
- This limitation is particularly challenging for clinical studies with scarce cell samples.
Purpose of the Study:
- To develop and optimize a combined assay for assessing multiple apoptosis stages.
- To enable analysis of apoptosis in limited clinical blood samples.
Main Methods:
- Developed two flow cytometry-based assays for peripheral blood mononuclear cells (PBMC).
- Assays measure caspase-12 activity, active caspase-3, DNA fragmentation, mitochondrial depolarization, and phosphatidylserine externalization.
- Validated apoptosis detection using ELISA.
Main Results:
- Optimized assays for typical clinical blood sample volumes and cell counts.
- The combined assays successfully differentiated apoptotic from non-apoptotic blood cells.
- Identified affected blood cell subsets within the samples.
Conclusions:
- A novel, combined two-assay method allows simultaneous assessment of major apoptosis pathways.
- This approach is suitable for analyzing apoptosis in clinical blood samples, even with limited cell numbers.
- Facilitates the study of apoptosis in relation to specific blood cell populations.
