Apoptosis Analysis

DNA degradation

At a late stage in the apoptotic cascade, a specific endonuclease breaks the DNA at the linkers between the nucleosomes to give large numbers of small fragments, whose sizes are 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. However, in apoptotic cells, 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 the intrinsic and/or extrinsic pathways. Caspases are destroying essential cellular proteins, leading to controlled cell death.

Flow Cytometry Facility
Barbara Pilas, Ph.D - Director
231 Edward R. Madigan Laboratory, 1201 W. Gregory Drive, Urbana, IL 61801
Phone: (217) 244-0559     FAX: (217) 244-0446      Email: pilas@uiuc.edu