How can apoptosis be measured

These panels include a collection of six knockout cell lines of genes involved in the apoptotic cell death pathway, all provided in one kit for your convenience, with matching parental cell lines as controls.

Annexin V binds to phosphatidylserine, which migrates to the outer plasma membrane in apoptosis. The analysis is typically done by flow cytometry. Pair Annexin V with a membrane-impermeable dye like 7-AAD to distinguish between intact, apoptotic, and necrotic cells eg see ab , ab , or ab See a partial list of our Annexin V dye conjugates below or a full list here.

Annexin V conjugate. Assay kits. DNA fragmentation can be measured using agarose gels. Gel electrophoresis. TUNEL assay. 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.

We also offer assays for cathepsin and calpain activity analysis: cathepsin D ab , cathepsin B ab , cathepsin L ab , and calpains ab 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. Necrosis, anoikis, and autophagy. We offer several kits for studying other forms of cell death: necrosis and apoptosis ab and ab , anoikis ab , and autophagy ab and ab Cell health assays guide.

Koopman, G. Blood 84 , PubMed Google Scholar. Collins, M. Elstein, K. Cell Res. Klucar, J. FEBS Lett. Fadok, V. Methods , 43— Gorczyca, W.

Gold, R. Green, D. Science , — Thornberry, N. Adiba Ishaque 1 Mohamed Al-Rubeai 2 3 1. Process Sciences Bayer Healthcare Berkeley 2. Personalised recommendations. While necrotic cell death is characterized by the release of endogenous 'danger signals' and subsequent inflammation, apoptosis is largely tolergenic. Therefore, care must be taken when assessing whether cells are dying via apoptosis or necrosis. Here, we highlight a number of assays, utilizing both microscopy and flow cytometry, to determine whether cells have undergone apoptosis or alternative modes of cell death.