STAUROSPORINE-INDUCED APOPTOSIS: ANALYSIS BY DIFFERENT ANNEXIN V ASSAYS
Jaromír Vašíček, Andrej Baláži, Peter Chrenek
Abstract
Jaromír Vašíček, Andrej Baláži, Peter Chrenek
Abstract
Annexin V assay is a well-known method for the evaluation of cell apoptosis. However, there is a wide range of available kits at the market that are of different quality and price. In this study, staurosporine-induced apoptosis model was used in order to compare low-cost and high-cost Annexin V detection kits from three different producers. Briefly, two human cell lines (KG-1 and NKT) were treated with staurosporine for 1, 3 and 6 hours and then evaluated for the apoptosis induction using 3 different detection kits by flow cytometry. Both cell lines showed significant (P<0.05) increase in cell apoptosis after 3 hours (20% and 13%, respectively) and 6 hours (50% and 20%, respectively) of incubation with staurosporine. No significant changes in the proportion of apoptotic cells were observed by comparing the data detected using low-cost or high-cost Annexin V assays. In conclusion, tested low-cost Annexin V detection kits could be safely used for an objective and fast flow cytometric assessment of the apoptosis in human or animal cells.
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Annexin V assay is a well-known method for the evaluation of cell apoptosis. However, there is a wide range of available kits at the market that are of different quality and price. In this study, staurosporine-induced apoptosis model was used in order to compare low-cost and high-cost Annexin V detection kits from three different producers. Briefly, two human cell lines (KG-1 and NKT) were treated with staurosporine for 1, 3 and 6 hours and then evaluated for the apoptosis induction using 3 different detection kits by flow cytometry. Both cell lines showed significant (P<0.05) increase in cell apoptosis after 3 hours (20% and 13%, respectively) and 6 hours (50% and 20%, respectively) of incubation with staurosporine. No significant changes in the proportion of apoptotic cells were observed by comparing the data detected using low-cost or high-cost Annexin V assays. In conclusion, tested low-cost Annexin V detection kits could be safely used for an objective and fast flow cytometric assessment of the apoptosis in human or animal cells.
Key concepts: Staurosporine, Annexin, Apoptosis, Flow cytometry, Annexin A5, Cell culture, Molecular biology, Programmed cell death