Cells were seeded at a density of 1 × 105 cells/well in a 12-well plate. At the 24-h point, cell morphology was assessed using the Apoptosis and Necrosis Assay Kit (Beyotime, Shanghai, China, Cat No. C0003). The cells were stained with Hoechst 33342 and propidium iodide (PI) according to the manufacturer’s protocols and then visualized by fluorescence microscopy. An Annexin-V-FITC/PI Cell Apoptosis Detection Kit (Meilunbio, Dalian, China, Cat No. MA0220-2) was used to detect the cells’ apoptotic rate according to the manufacturer’s direction. Afterward, the apoptotic rate of cells was measured within 1 h by a flow cytometer (BD Biosciences).
Annexin 5 fitc pi cell apoptosis detection kit
Manufactured by Meilun
Sourced in China
The Annexin-V-FITC/PI Cell Apoptosis Detection Kit is a laboratory tool used to detect and quantify apoptosis, a form of programmed cell death, in cell samples. The kit utilizes Annexin V, a protein that binds to phosphatidylserine, and propidium iodide (PI), a DNA-binding dye, to identify and differentiate between viable, early apoptotic, and late apoptotic/necrotic cells through flow cytometry or fluorescence microscopy.
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2 protocols using annexin 5 fitc pi cell apoptosis detection kit
1
Apoptosis and Necrosis Assessment
2
Annexin V-FITC/PI Apoptosis Assay
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Following incubation for 48 h, a AnnexinV-FITC/PI cell apoptosis detection kit (Meilunbio) was used to measure apoptotic cells. The cells (2x105) were centrifuged at 800 x g for 5 min at 4˚C, collected and washed with PBS twice at 4˚C. The cells (1x106/ml) were resuspended with 250 µl 1X binding buffer. The cells (100 µl) were subsequently added to a pipe with 5 µl Annexin V-FITC and 10 µl propidium iodide (PI; 20 µg/ml). The mixture was cultured at 25˚C for 15 min in darkness. The apoptotic rate was analyzed using a flow cytometer (Cytoflex3L8C; Beckman Coulter, Inc.) and the results were analyzed using Cell Quest software version 2.0 (BD Biosciences).
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