After transfection and OGD/R treatment, HL-1 cells were suspended in 195 ml of binding buffer at 5 × 105 cells/ml. Next, 5 ml of Annexin V-FITC solution (Sangon Biotech, Shanghai, China) was added, and the mixture was cultured for 15 min in the dark at room temperature. After centrifugation at ×·1000 g for 5 min and washing with binding buffer, cells were resuspended in the mixture of 190 ml of binding buffer and 10 ml of propidium iodide (PI; Sangon Biotech). Cells were further analyzed using flow cytometry (Beckman Coulter, Fullerton, CA, USA). This assay was performed for 3 times (n = 3).
Annexin 5 fitc solution
Manufactured by Sangon
Sourced in China
Annexin V-FITC solution is a fluorescently labeled protein that binds to phosphatidylserine, a phospholipid found on the surface of cells undergoing apoptosis or programmed cell death. This solution can be used to detect and quantify apoptotic cells in various experimental settings.
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Lab products found in correlation
3 protocols using annexin 5 fitc solution
1
Annexin V-FITC Apoptosis Assay
2
Annexin V-FITC Apoptosis Assay
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Cells were harvested at 48 h post-transfection and re-suspended in pre-cooled PBS. Next, 5 μL of Annexin V-FITC solution (Sangon Biotech, Shanghai, China) was added and incubated at room temperature in the dark for 15 min. After centrifugation and washing with binding buffer, cells were re-suspended in a solution containing 190 μL binding buffer and 10 μL propidium iodide (PI; Sangon Biotech). Then cell apoptosis was detected by flow cytometry on a Beckman flow cytometer (Beckman Coulter, Fullerton, USA).
3
Apoptosis Evaluation by Flow Cytometry
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Cells were harvested after 48 h for transfection, and re-suspended in the precooled phosphate buffered saline. 5 μl Annexin V-FITC solution (Sangon Biotech) was added to each well and incubated for 15 min at room temperature in the dark. After centrifugation and washed with binding buffer, cells were re-suspended again in solution containing 190 μl binding buffer and 10 μl propidium iodide (PI, Sangon Biotech). Then apoptosis rate was analyzed by using ow cytometry (Beckman Coulter, Fullerton, CA, USA).
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