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Detecting Cleaved Caspase-3 in Apoptotic Cells by Flow Cytometry
Lisa C Crowley1, Nigel J Waterhouse1,2,3
1Apoptosis and Cytotoxicity Laboratory, Mater Research, Translational Research Institute, Woolloongabba, Brisbane, Queensland 4102, Australia.
Cold Spring Harbor Protocols
|November 3, 2016
Summary
This study details a flow cytometry method to quantify apoptosis by detecting cleaved caspase-3. This approach reliably identifies cells undergoing or having undergone programmed cell death.
Area of Science:
- Cell Biology
- Biochemistry
Background:
- Apoptosis is a regulated cell death process orchestrated by caspases, a family of cysteine proteases.
- Active caspases cleave specific substrates, generating detectable fragments during apoptosis.
Purpose of the Study:
- To outline a protocol for quantifying apoptosis using flow cytometry.
- To highlight the detection of cleaved caspase-3 as a reliable marker for apoptotic cells.
Main Methods:
- Utilizing antibodies specific to caspase-cleaved fragments.
- Employing flow cytometry for the quantification of cells exhibiting caspase cleavage.
- Focusing on the detection of cleaved caspase-3, a key executioner caspase.
Main Results:
- Antibodies recognizing cleaved caspase fragments enable specific labeling of apoptotic cells.
- Flow cytometry allows for the quantification of these labeled cells.
- Detection of cleaved caspase-3 serves as a robust indicator of apoptosis.
Conclusions:
- Flow cytometry combined with cleaved caspase-3 detection offers a reliable method for quantifying apoptosis.
- This technique facilitates the identification and measurement of cells undergoing programmed cell death.
