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You can determine how cells are dying by measuring markers that are activated in different types, and at different stages, of cell death.
Annexin V binds to phosphatidylserine, which migrates to the outer plasma membrane in apoptosis. Analysis is typically by flow cytometry. Pair Annexin V with a membrane impermeable dye like 7-AAD to distinguish between intact, apoptotic, and necrotic cells (eg see ab214663, ab214484, or ab214485).
See a partial list of our Annexin V dye conjugates below or a full list here.
Annexin V conjugate
DNA condensation in apoptosis can be measured using DNA stains to visualize condensed nuclei.
DNA fragmentation can be measured using agarose gels. In the TUNEL assay, the 3’ ends of DNA fragments are labeled with deoxyuridine either conjugated to a fluorescent dye or biotin.
Flow cytometry, fluorescence microscope, microscope
Activated caspases can be detected using antibodies with IHC, western blotting, or flow cytometry.
Caspase activity assays either use peptide substrates, which are cleaved by caspases in cell extracts, or similar substrates that bind to activated caspases in live cells. Caspase specificity varies by substrate.
Learn more about our assays for caspases 1 through 12, formulated either for cell lysates with analysis by plate reader or for live cells with analysis by flow cytometer, microscope or plate reader.
Dyes that accumulate in mitochondria due to the mitochondrial membrane potential are also used in the analysis of apoptosis. For more information, see our guide to cell viability assays. Apoptotic cells stain more weakly with these dyes due to the loss of membrane potential.
Cytochrome C is released into the cytoplasm following total loss of mitochondrial membrane potential.
Western blot, fluorescence microscope
Glutathione assays are also used for the analysis of apoptosis.
Fluorometric plate reader
Learn more about:
- Cell viability assays based on cell metabolism/enzyme activity
Measure the rate of continuing cellular activities, such as metabolism.
- Cytotoxicity assays based on cytolysis/membrane leakage
Test for cell membrane damage, either by measuring the leakage of cellular enzymes or staining with membrane-impermeable dyes.
- Cell proliferation and cell cycle assays
Monitor the growth of a cell population using cell staining dyes, detect generations of daughter cells, or analyze the cell cycle state of a cell population.