The apoptotic rate of the chondrocytes was detected with Annexin V/PI staining (Nanjing KeyGen Biotech, Nanjing, China) using a fluorescence-activated cell sorting (FACS) machine (FACSCalibur™; Becton-Dicskinson Biosciences, San Diego, CA, USA). The changes in mitochondrial membrane potential (ΔΨm) were measured using a JC-1 kit (Nanjing Keygen Biotech), and the processed cells were also analyzed using the FACS machine. Both staining procedures were carried out according to the relevant manufacturer's instructions.
Annexin 5 pi staining
Manufactured by Keygen Biotech
Sourced in China
Annexin V/PI staining is a laboratory technique used to detect and quantify apoptosis (programmed cell death) in cell samples. It utilizes Annexin V, a protein that binds to phosphatidylserine (PS), and propidium iodide (PI), a DNA-binding dye, to differentiate between viable, early apoptotic, and late apoptotic/necrotic cells.
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6 protocols using annexin 5 pi staining
1
Apoptosis and Mitochondrial Membrane Potential
2
Berberine and Paclitaxel Combination Therapy
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The chemicals including berberine and paclitaxel were offered by Meilunbio. The purity of berberine was more than 99.8%. 2-Deoxy-D-glucose (2-DG) was purchased from Meck. The apoptosis detection kit (Annexin V-PI Staining) was purchased from Keygen Biotech (Nanjing, China). The Diff-Stain Set was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).
3
Annexin V-PI Apoptosis Assay
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Cell apoptosis was measured by Annexin V/PI staining (KeyGen Biotech, China) as described previously [15 (link), 39 (link)]. The cells were treated, harvested, and resuspended in 500 μl of binding buffer, and 5 μl of Annexin V and 5 μl of PI were added. Cell apoptosis was analyzed using flow cytometry.
4
Apoptosis Detection in H9c2 Cells
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Apoptosis levels of H9c2 cells were detected by Annexin V/PI staining (Cat: KGA108-2; Keygen; China). Briefly, cells were first digested using trypsin and collected at a concentration of approximately 1*10^6 cells/ml. Then, the cells were resuspended in a 200 µl binding buffer and then stained with 5 µl Annexin-FITC and PI. The Flow cytometer (BD; USA) and the FlowJo V10 software were used to analyze and calculate the apoptotic rate.
5
Annexin V Apoptosis Assay for Flow Cytometry
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Apoptosis of ALL cells was detected using an annexin V apoptosis assay, followed by flow cytometry analysis. In brief, cells were harvested following treatment, washed in PBS, and subjected to Annexin V/PI staining according to the manufacturer’s protocol (Keygen Biotech, Nanjing, China). The percentage of apoptotic cells was evaluated using flow cytometer (BD Accuri C6).
6
Quantifying Cell Death Mechanisms
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Annexin V-PI staining (KeyGen Biotech, China) was performed to measure the phosphatidylserine externalization in HepG2 cells transfected with different siRNAs or plvx vectors for the cell death detection. Briefly, trypsinized cells were collected, washed twice with ice-cold PBS, and resuspended in 200 μL binding buffer containing 5 μL FITC-conjugated Annexin V and 5 μL of propidium iodide. The staining sample was incubated at room temperature for 20 min, and 10,000 cells were immediately analysed using a FACSCalibur flow cytometer (BD Biosciences, USA).
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