Authors
Yingyu Cui
Summary
Apoptosis is a normal physiological phenomenon put forward by Kerr [1]. It plays an important role in embryonic development, maintenance of tissue homeostasis and pathology, and therefore has become a hotspot of activity in biomedical fields. To date, many methods to detect apoptosis have sprung up, including morphological identification, DNA ladder observation and DNA content and cell cycle analysis. However, these methods differ in treatment and are performed independently, consuming more time. As degraded low-molecular-weight DNA is exuded out of apoptotic cells after fixation with ethanol, DNA dyed with fluorescein will form an apoptosis curve (sub-diploid curve) because its contents are less than that of G1 phase [2]. The quantity of phosphate-citric acid buffer (0.2M, pH7.8) can effectively control the amount of the extracted DNA, and the exuded DNA can be used for DNA ladder detection through gel electrophoresis, while the residual cells can be analyzed with a flow cytometer [3,4]. Here I have uploaded an integrative procedure through which a relatively satisfactory result can be obtained following a single stage of cell culture and transient cell treatment, then detection with different instruments. This shortens experiment time. The protocols are as follows [5-7].Further details
The protocol was published on Protocol Exchange in 2006. To see the entire protocol, click on the source link.Advertisement
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