After the treatment, the CHON-001 cells were cultured in 96-well plates. The cells were then collected by centrifugation at 4 °C for 5 min. The cells were then washed twice with phosphate buffered saline and assessed using the Annexin V/propidium iodide apoptosis detection kit (Beyotime). The cells were gently mixed and incubated for 20 min at room temperature in the dark. Finally, the apoptotic cells were analyzed using a flow cytometer (Beckman Coulter, Brea, CA, USA) and Kaluza, according to the manufacturer’s instructions.
Annexin 5 propidium iodide apoptosis detection kit
Manufactured by Beyotime
Sourced in China
The Annexin V/propidium iodide (PI) apoptosis detection kit is a laboratory tool used for the detection and quantification of apoptosis, a type of programmed cell death. It utilizes the binding properties of Annexin V, a calcium-dependent phospholipid-binding protein, to detect the exposure of phosphatidylserine on the outer cell membrane, an early event in the apoptotic process. The kit also includes propidium iodide (PI), a fluorescent dye that stains the DNA of cells that have lost membrane integrity, indicating late-stage apoptosis or necrosis. The kit allows for the differentiation between early apoptotic, late apoptotic, and necrotic cells through flow cytometric analysis.
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Lab products found in correlation
Facscalibur flow cytometer, by BD (5 mentions)
Facscalibur, by BD (3 mentions)
Annexin 5 fitc, by BD (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Kaluza analysis software, by Beckman Coulter (1 mentions)
Facscanto 2, by BD (1 mentions)
Cellquest analysis software v 3, by BD (1 mentions)
Annexin 5 fitc, by Beyotime (1 mentions)
Cytomics fc500 flow cytometer, by Beckman Coulter (1 mentions)
Acea novocyte flow cytometer, by Agilent Technologies (1 mentions)
Crocin, by Merck Group (1 mentions)
Egfp lc3 plasmid, by Addgene (1 mentions)
Goat anti rabbit igg h l alexa fluor 594, by Abcam (1 mentions)
Z vad fmk, by Selleck Chemicals (1 mentions)
Cck 8, by Beyotime (1 mentions)
Flow cytometry, by Thermo Fisher Scientific (1 mentions)
Cell counting kit 8 cck 8, by Dojindo Laboratories (1 mentions)
Reactive oxygen species assay kit, by Beyotime (1 mentions)
Total superoxide dismutase assay kit with wst 8, by Beyotime (1 mentions)
Lipid peroxidation mda assay kit, by Beyotime (1 mentions)
23 protocols using annexin 5 propidium iodide apoptosis detection kit
1
Apoptosis Detection in CHON-001 Cells
2
Annexin-V/PI Apoptosis Assay in HeLa Cells
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After digesting the cells with trypsin without EDTA, the HeLa cells were collected by centrifugation at 4°C for 5 min. After that, the cells were washed twice with pre-cooled PBS. Then 1× binding buffer was used to prepare cell suspension. For cell apoptosis assay, cells were assessed using the Annexin-V/propidium iodide Apoptosis Detection Kit (Beyotime) according to the manufacturer’s protocol. Finally, apoptotic cells were checked using a Flow cytometer (BD Biosciences) and analyzed with Kaluza Analysis software (version 2.1.1.20653; Beckman Coulter, Inc.).
3
Annexin V/PI Apoptosis Assay
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Cell apoptosis was detected by the Annexin V/Propidium iodide (PI) Apoptosis detection kit (Biyuntian, Shanghai, China) according to the manufacturer’s protocol. Briefly, cells were grown in 6-well plate and then treated with indicated dosage of TN for 48h and collected. Cells were resuspended with 196μL binding buffer containing 5μL Annexin V and 10μL PI fluorescence dye and then apoptosis was detected by flow cytometry (Becton Dickinson FACS Calibur).
4
Apoptosis Detection Assay with Triptonide
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Annexin V/propidium iodide (PI) apoptosis detection kit (Biyuntian, Shanghai, China) was used to detect cell apoptosis according to the manufacturer’s protocol. CAL-27 cells were exposed to different concentrations of triptonide (10 nM, 30 nM, or 50 nM) for 72 h and then collected. Cells were resuspended in 200 μL binding buffer containing 5 μL Annexin V and 10 μL PI fluorescence dye, and then apoptosis was detected by flow cytometry (Becton Dickinson FACS Calibur).
5
Apoptosis Induction in Ovarian Cancer Cells
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A2780 and OVCAR-3 cells (1 × 106/well) were cultured in 6-well plates and treated with RC48, CM or their combination for 48 h. Flow cytometry was performed using the Annexin V-propidium iodide (PI) apoptosis detection kit (Beyotime Biotechnology, Shanghai, China) on a BD FACSCantoII instrument (USA) to determine cell apoptosis.
6
Annexin V-FITC and PI Apoptosis Assay
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Transfected cells were analyzed using the Annexin V/propidium iodide (PI) Apoptosis Detection Kit (Beyotime, Shanghai, China) [38 (link)]. In brief, we washed the cells twice with pre-cooled 1x phosphate-buffered saline before collecting the cells, and then prepared a cell suspension of 1 × 106 cells/mL using the fluorescein isothiocyanate (FITC)-binding buffer. We added 100 μL of the cell suspension to EP tubes. Subsequently, we added an appropriate amount of Annexin V-FITC and PI to the cells, according to the standard operating protocol. Cells were gently mixed and incubated for 20 min at 25°C in the dark. Cell apoptosis were analyzed using a BD FACSCalibur flow cytometer (BD Technologies). The data were analyzed using Kaluza Analysis (version 2.1.1.20653; Beckman Coulter, Inc.).
7
Annexin V-FITC Apoptosis Assay
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Cell apoptosis was performed by using the Annexin V/propidium iodide (PI) Apoptosis Detection Kit (Beyotime, Shanghai, China). Briefly, CAL-27 and SCC-9 cells were transfected with control plasmid or SCARA5 plasmid for 48 h and then the transfected cells were collected, disposed, and centrifuged at 1,000 × g at 4℃ for 5 min and re-suspended in 100 µL of FITC-binding buffer. Subsequently, we added approximately 5 µL of ready-to-use Annexin V-FITC and 5 µL of PI into the buffer, and the cells were incubated for 30 min at room temperature without light. Annexin V-FITC and PI fluorescence were assessed by BD FACSCalibur flow cytometer (BD Technologies).
8
Apoptosis Detection in Melanocytes
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Cell apoptosis detection was performed using the Annexin-V/propidium iodide (PI) Apoptosis Detection kit (Beyotime Institute of Biotechnology). Briefly, human melanocytes were plated into 6-well plates at a density of 2×105 cells per well. On the following day, the cells were transfected with inhibitor control, miR-421 inhibitor, miR-421 inhibitor + control-shRNA, or miR-421 inhibitor + RIPK1-shRNA. After transfection for 24 h, the cells were treated with 3 µM TM for 48 h. Subsequently, the cells were collected, centrifuged with low temperature at high speed (1,000 × g; 5 min; 4°C) and re-suspended in 100 µl fluorescein isothiocyanate (FITC)-binding buffer. Subsequently, ~5 µl ready-to-use Annexin V-FITC (BD Bioscience) and 5 µl PI were added to the buffer and the cells were incubated for 30 min at room temperature in the dark. Annexin V-FITC and PI fluorescence were assessed by BD FACSCalibur flow cytometer (BD Biosciences; Becton, Dickinson and Company). FlowJo software (version 7.6.1; FlowJo LLC) was used to analyze the data.
9
Annexin V-FITC Apoptosis Assay in Breast Cancer
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Apoptotic MCF-7 and MDA-MB 231 cells were investigated using the Annexin V/propidium iodide (PI) apoptosis detection kit (Beyotime Institute of Biotechnology) followed by flow cytometry, according to the manufacturer's protocol. MCF-7 and MDA-MB 231 cells (5×103 cells/well) were cultured in 6-well plates and treated with different concentrations (15, 30 and 60 µM) of Rosmanol for 48 h at 37°C. Cells were harvested by trypsinization with no EDTA and washed twice with PBS and then stained with 5 µl Annexin V-FITC (Beyotime Institute of Biotechnology) and 10 µl PI in 500 µl binding buffer for 15 min at room temperature (RT) in the dark. Apoptotic cells were determined by using Cytomics FC 500 flow cytometer (Beckman Coulter Inc.) and the data were analysed using Cellquest analysis software v.3.0 (BD Biosciences).
10
Apoptosis Detection via Annexin-V/PI Assay
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OGD-induced apoptosis was detected using an Annexin-V/propidium iodide (PI) apoptosis detection kit (Beyotime Institute of Biotechnology) according to the manufacturer's instructions. Briefly, N2A cells from each group were harvested after centrifugation at 90 × g for 5 min at room temperature and washed twice with PBS. Next, the cells were stained with Annexin V-FITC and PI for 15 min in the dark. Finally, apoptosis was analyzed using a flow cytometer (NovoCyte, Aceabio Biosciences, Inc.) and NovoExpress software version 1.3.1 (Aceabio Biosciences, Inc.).
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