After collection, digestion, and centrifugation, the cells were re-suspended in binding buffer (1×). Cell apoptosis was analyzed using the Annexin V-fluorescein isothiocyanate (FITC)/PI apoptosis detection kit (KeyGen, Jiangsu, China). Cell fluorescence was analyzed through a FACScan flow cytometry (Beckman Coulter, Brea, CA, USA).
Annexin 5 fluorescein isothiocyanate fitc pi apoptosis detection kit
Manufactured by Keygen Biotech
Sourced in China
The Annexin V-fluorescein isothiocyanate (FITC)/PI apoptosis detection kit is a laboratory instrument used to detect and quantify apoptosis (programmed cell death) in cell samples. It utilizes Annexin V, a protein that binds to phosphatidylserine, and propidium iodide (PI), a DNA-binding dye, to differentiate between viable, early apoptotic, and late apoptotic/necrotic cells.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Propidium iodide, by Merck Group (2 mentions)
Facscan, by Beckman Coulter (1 mentions)
Cyclin b1, by Abcam (1 mentions)
Penicillin, by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Caspase 3, by Abcam (1 mentions)
Cell cycle analysis kit, by Beyotime (1 mentions)
Facscanto 2, by BD (1 mentions)
Lsr 2, by BD (1 mentions)
Cell cycle analysis kit, by BD (1 mentions)
Celltrace cfse cell proliferation kit, by Thermo Fisher Scientific (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Rnase a, by Merck Group (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions)
Imatinib, by Novartis (1 mentions)
Facscalibur, by BD (1 mentions)
Propidium iodide, by Sangon (1 mentions)
Ethanol, by Beyotime (1 mentions)
Rnase a, by Sangon (1 mentions)
11 protocols using annexin 5 fluorescein isothiocyanate fitc pi apoptosis detection kit
1
Annexin V-FITC/PI Apoptosis Assay
2
Gastric and Hepatoma Cell Lines Characterization
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Human gastric cancer cell lines (SGC-7901 and MGC-803) and hepatoma cell lines (Bel-7402 and HepG2) were purchased from Sinochrome/Cytogenetics (Shanghai, China). All cell lines were cultured in Roswell Park Memorial Institute-1640 supplemented with 10% fetal bovine serum (Thermo Fisher Scientific, Waltham, MA, USA), 100 U/mL penicillin (Sigma-Aldrich Co., St Louis, MO, USA), and 100 μg/mL streptomycin (Sigma-Aldrich Co.) at 37°C in an atmosphere containing 5% CO2. To stably express Gluc and the cyan fluorescent protein (CFP), cells were transduced with the CSCW–Gluc–IRES–CFP lentivirus vector. rhCNB (enzyme unit < 0.25 U) was provided by Haikou Qili Pharmaceutical Co., Ltd (Hainan, China). The Annexin V-fluorescein isothiocyanate (FITC)/PI Apoptosis Detection Kit was purchased from KeyGen Biotech (Jiangsu, China). The Cell Cycle Analysis Kit was purchased from Beyotime (Shanghai, China). The following antibodies were used in this study: caspase-3, p53, cyclin B1 and CDK1 (purchased from Abcam, Cambridge, UK).
3
Annexin V-FITC/PI Apoptosis Assay
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Apoptosis assays were carried out using the Annexin V–fluorescein isothiocyanate (FITC)/PI apoptosis detection kit (Nanjing KeyGen Biotech, Inc.) according to the manufacturer’s protocol. Briefly, the cells were collected after transfection with siRNA for 48 h, washed twice with PBS, and resuspended in binding buffer. Sequentially, the cells were stained with Annexin V–FITC and PI at room temperature for 15 min and then analyzed by flow cytometry (BD, FACSCantoTM II).
4
Evaluating Apoptosis in ECA-109 Cells
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A total volume of 1×106 ECA-109 cells were treated with 0, 20, 40 or 80 mg/l COE for 24 h, and the harvested cells were washed twice with ice-cold PBS. Apoptotic cells were identified using the Annexin V-Fluorescein Isothiocyanate (FITC)/PI Apoptosis Detection kit (Nanjing KeyGen Biotech Co., Ltd., Nanjing, China). After centrifugation at 100 × g for 5 min at 4°C, 290 µl of 1X binding buffer, 5 µl of Annexin V-FITC and 5 µl of PI were added to the pellet, and incubated at room temperature (25°C) for 15 min in the dark. Next, 200 µl of 1X binding buffer was added prior to measurement. The data were measured and analyzed with the same machine and software as the cell cycle assay. The cell apoptosis assay was performed with 3 independent experiments.
5
Quantification of NPSC Apoptosis and Necrosis
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The Annexin V-fluorescein isothiocyanate (FITC)/PI Apoptosis Detection Kit (Nanjing Keygen Biotech), and the Cell Apoptosis Detection Kit (PI) (Nanjing Keygen Biotech) were used to quantify the apoptotic and necrotic ratio of NPSCs, respectively. Briefly, after compression treatment, cells were harvested by trypsinization and washed twice with PBS. Then, the cells were stained according to manufacturer’s instructions. All samples were subsequently analyzed by flow cytometry (BD LSR II, Becton Dickinson).
6
Apoptosis, Cell Cycle, and Proliferation
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Apoptosis and cell cycle were measured by the Annexin V-fluorescein isothiocyanate (FITC)/PI apoptosis detection kit (KeyGEN, Nanjing, China) and cell cycle analysis kit (BD Biosciences, SanJose, CA, USA) following the protocols, respectively. Cell proliferation was detected using CellTrace™ CFSE Cell Proliferation Kit (Thermofisher, USA). The results were analyzed with a FACSCalibur flow cytometer (BD Biosciences).
7
Cell Cycle and Apoptosis Analysis of DTX-Resistant Prostate Cancer Cells
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For cell cycle progression analysis, the transfected PC-3/DTX and DU145/DTX cells were seeded in dishes (6 cm) and disposed with 10 nM DTX. After about 80% confluence, the cells were collected and fixed with ethanol (75%). Next, the cells were stained with propidium iodide (PI) (500 μL, Sigma) in the presence of RNase A (20 μg/mL, Sigma). Thereafter, the cells were washed with phosphate buffer solution (PBS) and the distribution of the cells was determined with the FACScan flow cytometry (Gallios, Beckman, USA).
The Annexin V-fluorescein isothiocyanate (FITC)/PI apoptosis detection kit (KeyGen) was utilized for cell apoptosis assessment. In brief, the transfected PC-3/DTX and DU145/DTX cells were cultured for 48 h after 10 nM DTX treatment. Then, the cells (5 × 105) were resuspended in binding buffer (1×) and then stained with Annexin V-FITC (10 μL) and PI (5 μL). The samples were analyzed through the FACScan flow cytometry (Gallios).
The Annexin V-fluorescein isothiocyanate (FITC)/PI apoptosis detection kit (KeyGen) was utilized for cell apoptosis assessment. In brief, the transfected PC-3/DTX and DU145/DTX cells were cultured for 48 h after 10 nM DTX treatment. Then, the cells (5 × 105) were resuspended in binding buffer (1×) and then stained with Annexin V-FITC (10 μL) and PI (5 μL). The samples were analyzed through the FACScan flow cytometry (Gallios).
8
Resveratrol and Imatinib on K562 Cells
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K562 was purchased from Shanghai Bogoo Biotechnology. Co., Ltd. (Shanghai, China) and dissolved in dimethyl sulfoxide. Resveratrol was purchased from Hubei Jusheng Technology Co., Ltd. (Hubei, China). Imatinib was purchased from Novartis (Basel, Switzerland) .
Propidium iodide (PI) and 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The annexin Vfluorescein isothiocyanate (FITC)/PI apoptosis detection kit was purchased from KeyGEN Biotech (NanJing, China).
Propidium iodide (PI) and 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The annexin Vfluorescein isothiocyanate (FITC)/PI apoptosis detection kit was purchased from KeyGEN Biotech (NanJing, China).
9
Quantifying Nuclear DNA and Apoptosis
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To assess the distribution of nuclear DNA content, cells were treated with indicated concentrations of NexA for 48 h and fixed overnight in 75% ethanol at 4°C. Then cells were treated with 20 units/ml RNAase for 15 min at 37°C and stained with 50 µg/ml propidiumiodide (PI) prior to analysis using flow cytometry. For the determination of apoptotic cells, cells were treated with different doses of NexA for 48 h. The apoptotic cells were measured with an Annexin V-fluorescein isothiocyanate (FITC)/PI apoptosis detection kit (KeyGEN, Nanjing, China). The rate of apoptosis was detected by flow cytometry.
10
Quantifying Apoptosis in Bel-7402 Cells
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According to the protocol of the Annexin V-fluorescein isothiocyanate (FITC)/PI apoptosis detection kit (Nanjing KeyGen Biotech Co., Ltd., Nanjing, China) which contains binding buffer, annexin V-FITC and PI staining buffer, the collected Bel-7402 cells were resuspended in 500 µl binding buffer and sequentially mixed with 5 µl Annexin V-FITC and 5 µl PI. The mixture was incubated for 15 min at room temperature in the dark. Cell apoptosis was calculated with flow cytometry (FACSCalibur; BD Biosciences) and CellQuest 3.3 software.
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