Apoptosis Analysis
DNA degradation
At a late stage in the apoptotic cascade, a specific endonuclease breaks the DNA at the linkers between the nucleosomes resulting in a large number of small fragments (oligomers of about 180 bp). The DNA strand breaks may be detected by observation of a sub-G1 peak in the DNA histogram.
Annexin V
In normal viable cells, phosphatidylserine (PS) is located on the cytoplasmic surface of the cell membrane. In apoptotic cells however, PS is translocated from the inner to the outer leaflet of the plasma membrane thus exposing PS to the external cellular environment.
Changes in Mitochondrial Membrane Potential
DiOC6 is a green-fluorescent cationic dye that accumulates in active mitochondria and is useful in following changes in the membrane potential of the mitochondria that occur during programmed cell death.
Caspase Assay
In the early stages of apoptosis, cysteine-aspartic acid specific proteases called caspases are activated. The activation can occur through intrinsic and/or extrinsic pathways. Caspases destroy essential cellular proteins leading to controlled cell death.
Links
Fluorescence Spectra of Afformentioned Dyes
Annexin V - Alexa 488 conjugate
Vendors
Molecular Probes
Sigma
Software
FCS Express
Further reading
Flow Cytometry of Apoptosis (different methods: Rhodamine 123, Zenon Technology, FLICAs, Annexin V, PI, DAPI, TUNEL Assay etc.)
Protocol for staining dead cells with PI from Cancer Research Lab in UK
Protocol for PI staining (also cell cycle for adherent cells) from Arizona Research Labs
Apoptosis Analysis Guide from abcam
BDBiosciences Apoptosis Detection Kit
ThermoFisher Apoptosis Detection Kit
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