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Alexa Fluor 488 Annexin V/Dead cell apoptosis assay

VG Vinod Gangadevi
ST Sowjanya Thatikonda
VP Venkatesh Pooladanda
GD Geetanjali Devabattula
CG Chandraiah Godugu
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Dead cell apoptosis assay is quick and suitable method for identification of apoptotic cells as Annexin V, being phospholipid binding protein has more affinity towards phosphatidyl serine, which moves from inner plasma membrane leaflet to outer leaflet in apoptotic cells and thereby these apoptotic cells can be quantitatively estimated [40] For this assay, Alexa Fluor® 488 Annexin V/Dead cell apoptosis kit (Thermo Fischer Scientific, USA) was used. Initially, the assay was optimized by seeding HaCaT cells at cell density 4 × 104 and allowed to adhere, and further treated with SeNPs (0.5, 1 and 2.5 µg/ml) for 1 h followed by stimulation with EGF and incubated for 24 h. Then the assay was performed with aforementioned kit and followed the manufacturer protocol. Later the samples from all groups were analyzed by flow cytometry [41].

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