The apoptosis analysis was performed according to the manufacturer's protocol using the Annexin V/propidium iodide (PI) Apoptosis Detection Kit (BestBio, Shanghai, China). In brief, 3 × 10 5 cells were resuspended in 500 μL of binding buffer, stained with 5 μL Annexin V in the dark for 15 min at 4 ℃, and were incubated for another 5 min after 10 μL PI being added. The percentage of apoptotic cells was determined by using FACS flow cytometer equipped with the Cell Quest software (Beckman Coulter, CA, USA).
Annexin 5 propidium iodide apoptosis detection kit
Manufactured by BestBio
Sourced in China
The Annexin V/propidium iodide (PI) Apoptosis Detection Kit is a laboratory equipment used to detect and quantify apoptosis, a type of programmed cell death. The kit utilizes Annexin V, a protein that binds to phosphatidylserine, and propidium iodide, a fluorescent dye that stains DNA, to differentiate between viable, apoptotic, and necrotic cells.
Automatically generated - may contain errors
Lab products found in correlation
Cell quest software, by Beckman Coulter (1 mentions)
N acetylcysteine nac, by Selleck Chemicals (1 mentions)
Gallios flow cytometer, by Beckman Coulter (1 mentions)
Ferrostatin 1, by Selleck Chemicals (1 mentions)
Z vad fmk, by Selleck Chemicals (1 mentions)
Galios flow cytometer, by Beckman Coulter (1 mentions)
4 protocols using annexin 5 propidium iodide apoptosis detection kit
1
Annexin V/PI Apoptosis Assay Protocol
2
Apoptosis Induction by CDCA
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Apoptosis assay was detected by flow cytometry using Annexin V/propidium iodide (PI) apoptosis detection kit (BestBio Company, China). Cells were seeded into 6-well plates at the concentration of 5 × 105 cells/well for 24h, and treated with different concentrations of CDCA with or without pretreatment with N-acetylcysteine (NAC) (Selleck Chemicals, USA), SB203580 (MCE, China), Ferrostatin-1 (Selleck Chemicals, USA), or Z-VAD-FMK (Selleck Chemicals, USA) for 1h. Then the cells were collected and suspended with 300 μL binding buffer including 5 μL Annexin-V-FITC and 10 μL PI for 15min at 4 °C. The percentage of apoptotic cells was analyzed using a Gallios flow cytometer (Beckman Coulter, CA, USA).
3
Annexin V-PI Apoptosis Assay
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Apoptosis was assessed with an Annexin V/propidium iodide (PI) apoptosis detection kit (BestBio, Shanghai, China) according to the manufacturer’s protocol. Cells were harvested after different treatments and washed twice with PBS. Then, the cells were resuspended in 400 μL of binding buffer and stained with 5 μL of Annexin V for 15 min and 10 μL of PI for another 5 min in the dark at 4 °C. The percentages of apoptotic cells were analysed immediately using a Galios flow cytometer (Beckman Coulter, CA, USA).
4
Apoptosis Detection via Annexin V/PI Assay
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Annexin V/propidium iodide (PI) Apoptosis Detection Kit (BestBio, Shanghai, China) was used to detect apoptotic cells. Briefly, cells were cultured for 24, 48, and 72 hours at 37° in a 5% CO2 atmosphere. After incubation, adherent cells were detached with trypsin. These harvested cells were resuspended in complete RPMI 1640 medium and centrifuged at 1500 rpm for 5 minutes. Next, the cells were washed twice with ice-cold PBS and then stained simultaneously with Annexin V and PI, according to the manufacturer’s instructions, for 15 minutes in the dark. Finally, the binding buffer was added to each reaction tube, and the cells were analyzed by the flow cytometry method. Apoptosis was assayed by Annexin V/PI staining following manufacturer’s instructions. Briefly, the total apoptotic cells were made up of 2 parts: early apoptosis (Annexin V+ and PI– cells) and late apoptosis (Annexin V+ and PI+ cells). The data were analyzed by the WinMDI V2.9 software.
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