A549 cells with indicated treatment were harvested and fixed with 70% ethanol. These cells were then stained using propidium iodide (PI) and the cell cycle stage assessed by flow cytometry. To evaluate apoptosis, cells were cultured at 80% confluence and trypsinized and stained with a PI/Annexin V Apoptosis Detection Kit (Vazyme, Jiangsu Sheng, China). Data were collected and analyzed on a BD FACSC Flow Cytometer using FACSD software (BD Biosciences, San Jose, CA, USA).
Annexin 5 pi apoptosis detection kit
Manufactured by Vazyme
Sourced in China
The Annexin V-PI apoptosis detection kit is a laboratory product that can be used to detect and quantify apoptosis in cell samples. It utilizes Annexin V, a protein that binds to phosphatidylserine, and propidium iodide, a DNA-binding dye, to distinguish between viable, apoptotic, and necrotic cells.
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Lab products found in correlation
Facscalibur, by BD (2 mentions)
Facsc flow cytometer, by BD (1 mentions)
Facscan flow cytometer, by BD (1 mentions)
Eclipse e800 fluorescence microscope, by Nikon (1 mentions)
Calibur flow cytometer, by BD (1 mentions)
Cellquest software, by BD (1 mentions)
Tunel fitc apoptosis detection kit, by Vazyme (1 mentions)
Accuri c6 plus flow cytometer, by BD (1 mentions)
Sybr green polymerase chain reaction master mix, by Thermo Fisher Scientific (1 mentions)
Vitamin d3, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Primescript rt master mix, by Takara Bio (1 mentions)
Ascorbic acid, by Merck Group (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
β glycerophosphate, by Merck Group (1 mentions)
Cytexpert v2, by Beckman Coulter (1 mentions)
Cytoflex flow cytometer, by Beckman Coulter (1 mentions)
Cxp analysis software, by Beckman Coulter (1 mentions)
Fc500 cytometer, by Beckman Coulter (1 mentions)
14 protocols using annexin 5 pi apoptosis detection kit
1
Cell Cycle and Apoptosis Analysis
2
Cell Cycle and Apoptosis Analysis
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For cell cycle analysis, the drug-treated cells at 80% confluence were harvested and fixed with 70% ethanol. For apoptosis analysis, the cells were cultured in attachment and then trypsinized and stained with PI/Annexin V (Apoptosis Detection kit; Vazyme Biotech Co., Ltd., Nanjing, China). Data were collected and analyzed on a BD FACSCalibur™ flow cytometer and using BD FACS Loader software (BD Biosciences).
3
Cell Cycle and Apoptosis Analysis
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DDP-treated and control cells at 80% confluency were harvested and fixed with 70% ethanol. These were then stained using propidium iodide (PI) and the cell cycle stage was assessed by flow cytometry. To evaluate apoptosis, the cells were cultured at 80% confluency, trypsinized and stained with a PI/Annexin V Apoptosis Detection kit (Vazyme, Jiangsu Sheng, China). The data were collected and analyzed on a BD FACScan flow cytometer using FACS software (BD Biosciences, San Jose, CA, USA).
4
Assessing NK Cell Cytotoxicity
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Control or fucosylated NK cells and A20-luc+/yfp cells were co-cultured in 96-well plates at different effector/target (E: T) ratios. After co-cultured for 6h, apoptosis of yfp+ lymphoma cells were detected by Annexin V/PI Apoptosis Detection Kit (Vazyme, China). The cytotoxic activities of NK cells were represented by the apoptotic rates.
5
Apoptosis Assay in Cell Lines
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5 × 105 cells were seeded in 6-well plates per well overnight and treated with indicated drugs for 48 h. The collected total cells were stained with the Annexin V-PI apoptosis detection kit (Vazyme, China) and analyzed by flow cytometry (Becton Dickinson, USA). FlowJo 6.0 was used to analyze the results. Each assay was performed for three times.
6
Apoptosis Evaluation via Flow Cytometry
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Cells were performed by Annexin V/PI apoptosis detection kit (Vazyme, A211-02) according to the manufacturer’s manual. The apoptosis rate was measured by flow cytometer (FACS Calibur, BD).
7
Apoptosis Detection Protocols
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An Annexin V-PI apoptosis detection kit (Vazyme Biotech Co. Ltd., China) was used in a manner similar to that of a previous description [22 (link), 23 (link)] to detect apoptosis. Thereafter, fluorescence-activated cell sorting (FACS) was utilized to count the stained cells using CellQuest software (BD Biosciences, USA) connected to a Calibur flow cytometer. A TUNEL FITC Apoptosis Detection Kit (Vazyme Biotech Co. Ltd., China) was used, as instructed by the manufacturer, to conduct TUNEL staining. Immunofluorescence was observed using a Nikon ECLIPSE E800 fluorescence microscope.
8
Annexin V-FITC/PI Apoptosis Assay
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Cell apoptosis was detected using Annexin V/PI apoptosis detection Kit (Vazyme, China) according to the manufacturer's instructions. The cells were collected and washed twice after 36–48 h of treatment. Thereafter they were suspended in 400 μL of binding buffer, 10 μL of PI solution and 5 μL of Annexin V antibody conjugated with fluorescein isothiocyanate (FITC). After 15 min at 4 °C in a dark environment of incubation in the mixture, apoptosis rate was measured by flow cytometry. Apoptosis was quantified using FlowJo-V10 software (BD Biosciences, San Jose, CA, USA).
9
Apoptosis Analysis in Pancreatic Cancer Cells
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Apoptosis was analyzed using the Annexin V-PI apoptosis detection kit (A211, Vazyme Biotech Co., Ltd, Nanjing, China). Briefly, the treated BxPC-3 and PANC-1 cells were trypsinized and washed twice with ice-cold PBS and re-suspended in the binding buffer at a concentration of 105 per well in a total volume of 100μl. Then, the cells were supplied with 5 μl of Annexin V-FITC and 5 μl of PI and incubated in dark at room temperature for 10 min. After that, 200 μl of binding buffer was added into each sample for flow cytometry analysis.
10
Annexin V-PI Apoptosis Assay
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LLC cells were collected and stained with Annexin V and PI according to the instruction of Annexin V-PI Apoptosis Detection Kit (A211-01/02, Vazyme Biotec, Nanjing, China). The fluorescence intensity was detected by BD Accuri™ C6 Plus Flow Cytometer (660517, Becton, Dickinson and Company) and the data analysis was performed by Flowjo software (Tree Star, Ashland, OR, USA).
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