An Annexin V-FITC/PI assay was conducted using the Annexin V-FITC Apoptosis Detection kit (70-AP101-100; Liankebio, Hangzhou, China) according to the manufacturer's protocol, followed by flow cytometric analysis. Briefly, NRK-52E cells were collected, washed twice with cold PBS and then resuspended in binding buffer. The cells were subsequently incubated with 10 µl Annexin V-FITC and 5 µl PI for 5 min at room temperature in the dark, followed by analysis by flow cytometry.
Annexin 5 fitc apoptosis detection kit
Manufactured by Liankebio
Sourced in China
The Annexin V-FITC Apoptosis Detection Kit is a laboratory tool used to detect and quantify apoptosis, a type of programmed cell death. The kit utilizes Annexin V, a protein that binds to phosphatidylserine, a molecule that is exposed on the surface of cells undergoing apoptosis. The Annexin V is conjugated to the fluorescent dye FITC, allowing for the visualization and analysis of apoptotic cells via flow cytometry or fluorescence microscopy.
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4 protocols using annexin 5 fitc apoptosis detection kit
1
Annexin V-FITC/PI Apoptosis Assay
2
Annexin V-FITC Apoptosis Assay in HCC Cells
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HCC cells were grown to 80% confluence on six-well plates, after which serum-containing medium was replaced by medium without serum. After 24 h of cell incubation without serum, all cells were collected. Cell apoptosis was detected using Annexin V-FITC apoptosis detection kit (Lianke, Hangzhou, China). Briefly, HCC cells were stained with Annexin-V-FITC/Propidium Iodide (PI) followed by Cytoflex flow cytometer (Beckman, China) analysis within 5 min of staining. Data were analyzed using FlowJo software (version 10.6.0, Ashland, OR, USA).
3
Evaluating Antioxidant Compounds against Oxidative Stress
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PIG1 cells were cultured in 60 mm petri dishes after treatment with 50 μmol/L QU, LU, or KA for 24 h. Then, H2O2 (final concentration: 1 mmol/L) was added to each well, and incubation was undertaken for an additional 2 h. Simultaneously, we set up simple compound-treatment groups and a control group (without any treatment). After the previous treatment, we digested PIG1 cells with EDTA-free trypsin, collected them in tubes, washed them twice with phosphate-buffered saline (PBS), and resuspended them in PBS. According to the protocol, PIG1 cells were stained with an Annexin V-FITC Apoptosis Detection Kit (Liankebio, Hangzhou, China) and detected by flow cytometry. FlowJo (http://www.flowjo.com/ ) was used to measure the percentage of apoptotic cells.
4
Annexin V-FITC Apoptosis Detection
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Cell apoptosis was detected using Annexin V-FITC Apoptosis Detection Kit (LiankeBio, China) according to the manufacturer's protocol [38 (link)]. 1 × 106 ~ 3 × 106 cells were harvested and washed with cold PBS. Subsequently, the cells were incubated with 5 μl of Annexin V-FITC and 10 μl of propidium iodide at room temperature for 5 min. Samples were measured using flow cytometry immediately.
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