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Measuring multiple apoptosis,,,parameters with the Agilent 2100,,,bioanalyzer

used by nature during development6, homeostasis, aging and in defense mechanisms. Apoptosis is a key factor in cancer and also suspected to be a characteristic mechanism for degenerative disorders such as Alzheimer7 or Parkinson's diseases. The process is universal and has been described in eukaryotic cells as well as in individual bacteria8, protozoa9 and amoeba10. It is a target for research scientists in a wide variety of fields. Apoptosis is characterized by a distinct set of morphological events involving plasma membrane blebbing and asymmetry loss, reduction of cell volume, loss of mitochondrial membrane potential, nuclear condensation, fragmentation of DNA at nucleosomal intervals and other cytoplasmatic changes. Ultimately fragmentation of the cell into membrane-enclosed apoptotic bodies occurs. Differential expression patterns or alternate mechanisms in the apoptosis pathways among tissues and cell lines makes it necessary to have two or even three parallel experiments to confirm, quantitate and compare kinetic events on apoptosis. The appearance of DNA laddering is unambiguously connected to apoptosis. Targeting of phosphatidyl serine (PS) in the outer leaflet of cell membrane with fluorescently labeled annexin V and antibody labeling of active-caspase- 3 in cells has been previously described in conjunction with the Agilent 2100 bioanalyzer as a quantitative measurement for apoptotic cell samples5. This Application Note describes the combined measurement of annexin V and the extracted DNA of apoptotic cells on one platform the Agilent 2100 bioanalyzer. A fast protocol for DNA preparation and recommendations for data evaluation are discussed.
Experimental
Apoptosis was induced in Jurkat cells by incubation in a 5 M solution of the topoisomerase inhibitor Camptothecin
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