H22 and BEL-7404 cells were treated with different concentrations of AAEE, AAEE-Pe, and AAEE-Ea for 24 h, and then stained with an Annexin V-FITC/propidium iodide (PI) Apoptosis Detection Kit (YEASEN, Shanghai, China) according to the manufacturer’s instructions. Cisplatin and DMSO were used as positive and negative controls, respectively. Samples were analyzed by flow cytometry (BD FACSCalibur, San Jose, CA, USA). This experiment was conducted three times independently.
Annexin 5 fitc propidium iodide apoptosis detection kit
Manufactured by Yeasen
Sourced in China
The Annexin V-FITC/propidium iodide (PI) Apoptosis Detection Kit is a laboratory tool used to detect and quantify apoptosis, a form of programmed cell death. The kit utilizes Annexin V, a protein that binds to phosphatidylserine, and the dye propidium iodide, which stains the nucleic acids of cells with compromised membranes. This combination allows for the identification and differentiation of viable, early apoptotic, and late apoptotic/necrotic cells.
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Lab products found in correlation
Facscalibur, by BD (2 mentions)
Flow cytometry, by BD (1 mentions)
Calcein am pi, by Solarbio (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Beyotime (1 mentions)
Cm5 sensor chip, by GE Healthcare (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Coumarin 6, by Meilun (1 mentions)
Sirna mate, by GenePharma (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Meilun (1 mentions)
Phosphatase inhibitor cocktail, by Yeasen (1 mentions)
Cholesterol, by Sinopharm (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Meilun (1 mentions)
Facscan, by BD (1 mentions)
Accuri c6, by BD (1 mentions)
Phosphate buffered saline (pbs), by Solarbio (1 mentions)
10 protocols using annexin 5 fitc propidium iodide apoptosis detection kit
1
Apoptosis Analysis of AAEE Compounds
2
Apoptosis and Cell Cycle Analysis
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flow cytometry was employed to detect cell apoptosis rate and cell cycle. Referring to manufacturer’s instruction, Annexin V-FITC/propidium iodide (PI) Apoptosis Detection Kit (YEASEN) was selected for evaluation of cell apoptosis rate. About 5 μL of Annexin V-FITC (labelled as early apoptotic cells) and 10 μL of PI (labelled late apoptotic and necrotic cells) Staining Solution were added in the cell suspension for 15 min.
For the detection of cell cycle, cells were fixed with ethanol for 48 h at 4°C and subsequently stained with PI. Finally, cell apoptosis rate and cell cycle were evaluated by flow cytometry (BD, NJ, USA).
For the detection of cell cycle, cells were fixed with ethanol for 48 h at 4°C and subsequently stained with PI. Finally, cell apoptosis rate and cell cycle were evaluated by flow cytometry (BD, NJ, USA).
3
Apoptosis Induction in H22 Cells
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H22 cells were treated with different concentrations of CTPG (0, 100, 200, 300 and 400 μg/ml) or 0.3% DMSO for 24 h, and then stained with Annexin V-FITC/Propidium iodide (PI) Apoptosis Detection Kit (YEASEN, China) according to the manufacturer’s instructions. Samples were analyzed by flow cytometry (BD FACSCalibur, USA).
4
Cytotoxicity and Apoptosis Evaluation
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All chemicals were of analytical grade and used directly without further purification. Polyvinylpyrrolidone (PVP, K30), potassium ferricyanide (K3[Fe (CN)6] 98%), and hydrochloric acid (HCl, 36.0%~38.0%) were purchased from the Yongsheng Fine Chemicals Co., LTD (Tianjin, China). PC B2 was purchased from Sunny Biotech Co., LTD (Shanghai, China). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was obtained from Beyotime Biotechnology Co. (Haimen, China). Annexin V-FITC/propidium iodide (PI) apoptosis detection kit was supplied by Yeasen Biotech Co., Ltd. (Shanghai, China). Calcein-AM/PI was purchased from Solarbio Science & Technology Co., LTD (Beijing, China). Phosphate buffer solution (PBS) and RPMI 1640 medium were obtained from GIBCO/Invitrogen Co., LTD (Beijing, China). Fetal bovine serum (FBS) was purchased from Biological Industries Co., LTD (Israel).
5
Evaluating Nanoxel-PM Delivery Efficacy
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DTX, ginsenoside 20 (S)–Rg3, and Nanoxel-PM (Samyang Biopharm) were gifted by Bensu Medicine Technology Co. Ltd. (Shanghai, China). Cholesterol was purchased from Sinopharm Chemical Reagent Co. Ltd. (Shanghai, China). PL-100M was purchased from AVT Medicine Technology Co. Ltd. (Shanghai, China). Coumarin-6 (C6), Hoechst 3334w, DAPI, 1′-dioctadecyl-3,3,3′,3′-tetramethylindotricarbocyanine iodide (DiR), DiD, MTT, and 1% crystal violet solution were purchased from Meilun Biotechnology Co., China. Annexin V–FITC/propidium iodide (PI) apoptosis detection kit and phosphatase inhibitor cocktail were provided by Yeasen Biotechnology Co., China. Penicillin, streptomycin, and 0.25% trypsin-EDTA were purchased from Invitrogen Co., USA. Protein GLUT1 was obtained from Wuhao Bio-Tech Co., China. CM5 sensor chips were purchased from GE Healthcare Bio-Sciences AB (Sweden). Anti-Glut1 small interfering RNA (siRNA) (CCAACUGGACCUCAAACUUTT, AAGUUUGAGGUCCAGU-UGGTT) and siRNA-mate were provided by GenePharma Co., China. All other chemicals and solvents used in this study were of reagent grade or high-performance liquid chromatography (HPLC) grade.
6
Apoptotic and Cell Cycle Analysis of BRBS-Treated A549 Cells
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A549 cells were treated with different concentrations (200, 400 and 600 μg/mL) of BRBS. After 24 h, cells were harvested, washed with phosphate-buffered saline (PBS) and stained with an Annexin V-FITC/propidium iodide (PI) Apoptosis Detection Kit (YEASEN, Shanghai, China) according to manufacturer’s instructions. For analysis of the cell cycle distribution, cells were harvested after BRBS treatment and fixed in cold 70% ethanol at 4 °C for 30 min. Then, cells were stained with PI for 30 min at 37 °C. All samples were analyzed by flow cytometry.
7
Apoptosis analysis of PC9/GR cells
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PC9/GR cells were seeded overnight in 6‐well plates at 1 × 105 cells/well. They were then treated with DMSO, 0.1 μM gefitinib, 2 μM anlotinib, or a combination of 0.1 μM gefitinib and 2 μM anlotinib. Cell apoptosis detection was performed using Annexin V‐FITC/propidium iodide (PI) Apoptosis Detection Kit (40302, Yeasen) according to the manufacturer's instructions. The treated cells were briefly harvested and washed with phosphate‐buffered saline, followed by suspension in 100 μl of Annexin V binding buffer. Then, 5 μl of Annexin V‐FITC and 10 μl of PI were added for incubation in the dark. Samples were then analyzed using a flow cytometer (FACScan, BD Biosciences). Experiments were performed in triplicate, and an average was obtained.
8
Apoptosis Quantification with Annexin V-FITC/PI
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Apoptosis abilities were detected by an Annexin V-FITC/Propidium Iodide (PI) Apoptosis Detection Kit (Yeason, China) on the basis of the manufacturer's protocol. The results were obtained using FlowJo software.
9
Osteoclast Progenitor Apoptosis Analysis
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The effects of diluted solutions of powder extracts on the apoptosis of osteoclast progenitors were analyzed using an Annexin V-FITC/propidium iodide (PI) apoptosis detection kit (403302ES20, Yeasen Biotechnology). According to the manufacturers’ instructions, cells were seeded in 6-well culture plates at a density of 4 × 105 per well. After an incubation period of 24 h, cells in diluted solutions (dilution ratios: 1/4, 1/8, 1/16, 1/32, 1/64 and 1/128) of BG-/Mo-extracts were digested, centrifuged, rinsed, and incubated in 200 µL of PBS containing 5 μL of Annexin V-FITC and 5 μL of PI for 15 min at room temperature. Subsequently, all the obtained cells were rinsed and subjected to flow cytometry (BD Accuri C6, San Jose, CA, USA) to measure the apoptosis rate (%).
10
Apoptosis Evaluation via Flow Cytometry
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The flow cytometry was performed using the Annexin V-FITC/propidium iodide (PI) Apoptosis Detection Kit (Yeasen, Shanghai, China). Cold phosphate-buffered saline (PBS) (Solarbio, Beijing, China) was used for washing. After washing twice with cold PBS buffer, the CRC cells were centrifuged at 4°C and 5 min later and made into 1 × 106 cells/mL suspension with 100 L of 1× binding buffer. After 15 min of complete reaction with 10 μL of PI solution and 5 μL of Annexin V-FITC, the mixture was placed on ice for detection by flow cytometry (BD Biosciences, USA).
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