Apoptotic cells were quantitated with an Annexin V-FITC/PI apoptosis detection kit (Multi Sciences Biotech, AP101-100), as directed by the manufacturer. In brief, after staining of H9c2 cells with Annexin V and PI for 30 min at ambient, a FACScan flow cytometer (BD FACSAria, USA) was utilized for analysis within 1 h.
Annexin 5 fitc pi apoptosis detection kit
Manufactured by MultiSciences Biotech
Sourced in China, United States
The Annexin V-FITC/PI Apoptosis Detection Kit is a laboratory product used for the detection and analysis of apoptosis in cell samples. It contains Annexin V-FITC, a fluorescent dye that binds to phosphatidylserine, and propidium iodide, a DNA-binding dye. The kit provides a method for identifying and quantifying apoptotic and necrotic cells through flow cytometry or fluorescence microscopy.
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Lab products found in correlation
Cell cycle staining kit, by MultiSciences Biotech (4 mentions)
Facscalibur ow cytometer, by BD (3 mentions)
Facscalibur flow cytometer, by BD (2 mentions)
Cellquest software, by BD (2 mentions)
Facscan flow cytometer, by BD (1 mentions)
Epics altra 2, by Beckman Coulter (1 mentions)
Flow cytometry, by BD (1 mentions)
Facsaria 3 cell sorter, by BD (1 mentions)
Annexin 5 apc pi apoptosis detection kit, by MultiSciences Biotech (1 mentions)
Cytoflex s, by Beckman Coulter (1 mentions)
Cytoflex system, by Beckman Coulter (1 mentions)
Facscanto 2, by BD (1 mentions)
Facsdiva software, by BD (1 mentions)
Dna staining solution, by MultiSciences Biotech (1 mentions)
Permeabilization solution, by MultiSciences Biotech (1 mentions)
Facsverse, by BD (1 mentions)
Dulbecco s modified eagle s medium f12 dmem f12, by Thermo Fisher Scientific (1 mentions)
3 methyladenine 3 ma, by Merck Group (1 mentions)
Gapdh antibody, by Abcam (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
27 protocols using annexin 5 fitc pi apoptosis detection kit
1
Annexin V-FITC/PI Apoptosis Assay
2
Quantifying Apoptosis with Annexin V-FITC/PI
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Annexin V-FITC/PI apoptosis detection kit (Multisciences) was used to analyze apoptosis. HK1-EBV and HONE1 cells were incubated with 0.006% DMSO (vehicle control) or curcumin (5 μM and 10 μM). After 24 h, cells were resuspended in the binding buffer (500 μl). The cells were incubated with Annexin V-FITC (5 μl) and PI (10 μl). After that, cells were analyzed with EPICS Altra II (Beckman Coulter).
3
Apoptosis Evaluation using Flow Cytometry
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After treatment, cells were harvested and stained with an Annexin V-FITC/PI Apoptosis Detection Kit (MultiSciences) according to the manufacturer's guide. The apoptosis was evaluated using flowcytometry (BD Bioscience), and data were analyzed using FlowJo V10.
4
Annexin V-FITC/PI Apoptosis Assay
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Apoptosis was detected using an annexin V-FITC/PI apoptosis detection kit or annexin V-APC/PI apoptosis detection kit according to the manufacturer's instructions [MultiSciences (Lianke) Biotech Co., Ltd.]. Briefly, VSMCs were plated into six-well plates and incubated with drugs for optimal intervals using the results from MTT assay. The cells were harvested, washed twice with pre-chilled PBS, and centrifuged (1,000 x g for 5 min at 4˚C), followed by staining with annexin V-FITC/PI or annexin V-APC/PI at 4˚C for 10 min in the dark and then stained cells were analyzed by flow cytometry (BD FACSAria™ III Cell Sorter; cat. no. 648282; BD Biosciences). Flow cytometry data analysis of the proportion of apoptotic cells was performed using the FlowJo 7.6.1 software (Flowjo LLC).
5
Cell Cycle and Apoptosis Analysis
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Cell cycle analysis was performed using the Cell Cycle Staining kit (70-CCS012, Multi Sciences, Hangzhou, China) according to the manufacturer’s protocol. For the cell apoptosis analysis, the Annexin V-FITC/PI Apoptosis Detection Kit (70-AP101–100, Multi Sciences) was used according to the manufacturer’s instructions. The stained cells were washed with PBS and analyzed via flow cytometry on a CytoFLEX System (CytoFLEX S, Beckman Coulter, Brea, California, USA).
6
Quantifying Apoptosis and Cell Cycle
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The Annexin V-FITC/PI Apoptosis Detection Kit (Multi Sciences Biotech Co., Ltd.) was used in the present study. Briefly, cells were seeded into 6-well plates and transfected. Cells were harvested and resuspended in 500 ul binding buffer after 48 h. Subsequently, cells were incubated with 5 μl Annexin V-FITC and 10 μl PI at room temperature in the dark for 15 min before analysis via flow cytometry.
For the cell cycle assay, the Cell Cycle Staining Kit (Multi Sciences Biotech Co., Ltd.) was used according to the manufacturer's protocol. Briefly, cells were harvested and fixed in 70% ethanol at 4 °C overnight. Cells were then treated with 100 μl RNase A at 37 °C for 30 min and incubated with 400 μl PI at 4 °C in the dark for 30 min. Cells were then analyzed via flow cytometry.
For the cell cycle assay, the Cell Cycle Staining Kit (Multi Sciences Biotech Co., Ltd.) was used according to the manufacturer's protocol. Briefly, cells were harvested and fixed in 70% ethanol at 4 °C overnight. Cells were then treated with 100 μl RNase A at 37 °C for 30 min and incubated with 400 μl PI at 4 °C in the dark for 30 min. Cells were then analyzed via flow cytometry.
7
Annexin V-FITC/PI Apoptosis Assay
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The Annexin V-FITC/PI Apoptosis Detection Kit (Multi Sciences Biotech Co., Ltd.) was used in the present study. Briefly, cells were seeded into 6-well plates and were harvested and resuspended in 500 μl binding buffer after 48 h. Subsequently, cells were incubated with 5 μl Annexin V-FITC and 10 μl PI at room temperature in the dark for 15 min before analysis via flow cytometry.
8
Apoptosis Induction in HFLS-RA Cells
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HFLS-RA cells were seeded into 6-well plates at the density of 1×10 5 / well and incubated for 12 h. The cells were exposed to different concentrations of FLA for 1 hour and then treated with IL-1β for another 24 hours. By then, the cells were collected by centrifugation at 800 × g for 10 minutes, and washed three times with precooled phosphate buffer saline (PBS). According to the introduction of the Annexin V-FITC/PI Apoptosis Detection Kit (Multi Sciences, Hangzhou, China), the cells were resuspended with 500 µL of binding buffer and incubated with 5 µL of Annexin V-FITC and 10 µL of PI for 5 minutes at room temperature in the dark. Apoptotic cells were detected by utilizing a FACSCalibur ow cytometer (BD Biosciences, San Jose, CA).
9
ACSS2 Regulates Cell Cycle and Apoptosis
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The effect of ACSS2 on cell cycle distribution was determined by flow cytometry. Briefly, cells subjected to the indicated treatments were harvested when they reached 80% confluence, washed with PBS and resuspended in 1 ml of DNA staining solution and 10 µl of permeabilization solution (MultiSciences Biotech Co., Ltd., Hangzhou, China). Following incubation for 30 min in the dark at room temperature, the cells were analyzed by flow cytometry at 72 h after interference. The fractions of cells in the G0/G1, S, and G2/M phases were analyzed. Cell apoptosis was detected using an Annexin V-FITC/PI Apoptosis Detection kit (MultiSciences Biotech Co., Ltd., Hangzhou, China) according to the supplier's protocol. Cells were seeded in 12-well plates (6×104/well). The cells were harvested after transfection with siRNA or NC and with or without DDP (5 µg/ml) treatment for 24 h, and washed in cold PBS. Then, cells were resuspended in 500 µl of binding buffer. Next, 5 µl of Annexin V-FITC and 10 µl of PI working solution were added to each reaction system at room temperature for 5 min. Flow cytometric analysis was performed immediately on a flow cytometer (BD FACSCanto II; BD Biosciences) and data were analyzed with BD FACSDiva software (BD Biosciences). Each experiment was repeated at least three times.
10
Apoptosis Induction in HFLS-RA Cells
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HFLS-RA cells were seeded into 6-well plates at the density of 1×10 5 / well and incubated for 12 h. The cells were exposed to different concentrations of FLA for 1 hour and then treated with IL-1β for another 24 hours. By then, the cells were collected by centrifugation at 800 × g for 10 minutes, and washed three times with precooled phosphate buffer saline (PBS). According to the introduction of the Annexin V-FITC/PI Apoptosis Detection Kit (Multi Sciences, Hangzhou, China), the cells were resuspended with 500 µL of binding buffer and incubated with 5 µL of Annexin V-FITC and 10 µL of PI for 5 minutes at room temperature in the dark. Apoptotic cells were detected by utilizing a FACSCalibur ow cytometer (BD Biosciences, San Jose, CA).
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