Kinetic Apoptosis and Necrosis Analysis of HT1080 cells

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HT1080 cells were plated at a density of 2000 cells/well in 96 well plates and allowed to adhere overnight. The next day, cells were treated with dilutions of camptothecin, starting at 2500 nM, in order to determine the effect of camptothecin on the apoptotic and necrotic response of HT1080 cells. Our results show an increase over time in the percentage of GFP positive apoptotic cells (A) and PI positive necrotic cells (B). Apoptotic and necrotic cells are normalized to high contrast bright field cell counts. This increase is dependent upon concentrations of camptothecin. The accompanying figure also shows a dose response curve for apoptosis (C) and necrosis (D), demonstrating that both the apoptotic and necrotic response show a dose dependent increase following treatment with camptothecin. The dose response curve is based on 48 hours of camptothecin treatment.
This kit was straightforward and easy to use. We performed the assay according to the manufacturer’s instructions using the lowest recommended doses of PI and pSIVA (5 and 10 ul/ml respectively). This assay was performed on a Cytation 5 Cell Imaging Multi-Mode Reader in conjunction with a BioSpa 8 Automated Incubator (BioTek Instruments, INC.).

Dr. Sarah Beckman

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Submitted Sep 18 2017

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