ATC cells were cultured with compounds for 48 h. Cultured cells were harvested using trypsin, washed with phosphate‐buffered saline, and fixed in 70% ethanol for more than 12 h at −20°C. The cells were then stained with propidium iodide solution. Cell cycle was analyzed using a flow cytometer FC500 (Beckman Coulter, CA, USA) and the MultiCycle for Windows cell cycle analysis software (Beckman Coulter, CA, USA).
Multicycle for windows
Manufactured by Beckman Coulter
Sourced in United States
Multicycle for Windows is a software program that provides automated cell cycle analysis. It offers tools for data acquisition, modeling, and reporting of cell cycle data obtained from flow cytometry experiments.
Automatically generated - may contain errors
Lab products found in correlation
Fc500 flow cytometry system with cxp software, by Beckman Coulter (2 mentions)
Fc500 flow cytometer, by Beckman Coulter (1 mentions)
Annexin 5 fitc pi staining kit, by BD (1 mentions)
Epics xl, by Beckman Coulter (1 mentions)
Annexinv labeled protein, by Beyotime (1 mentions)
Coulter dna prep reagents kit, by Beckman Coulter (1 mentions)
5 protocols using multicycle for windows
1
Cell Cycle Analysis of ATC Cells
2
Apoptosis Analysis in H1299 Cells
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H1299 cells were transfected with TFAP2B siRNA. At 48 h after transfection, the cells were harvested by trypsinization and fixed in 70% cold ethanol for 30 minutes, then stained with 5 μl Annexin V-FITC and 5 μl PI (propidium iodide) using an Annexin V-FITC/PI-staining kit (Becton Dickinson, CA, USA). The cells were placed at room temperature for 15 min in the dark and then analyzed using flow cytometry (EPICS XL; Beckman Coulter). Apoptosis was calculated in terms of the FITC-positive cells, and the PI staining was used to perform a cell cycle analysis. The raw data were analyzed using Multicycle for Windows (Beckman Coulter).
3
Apoptosis Evaluation of Ovarian Cells
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After transfection of ovarian granulosa cells for 48 h, the cells were detached with trypsin for 3 min, and collected in a 10 mL centrifuge tube. Then, cells were fixed overnight with pre-cooled 70% ethanol, AnnexinV labeled protein (Beyotime Institute of Biotechnology, Shanghai, China) and corresponding nucleic acid dye was added, evenly mixed and reacted for 15 min. Binding buffer (1 × 400 μL) was put into the flow tube, and the DNA data of the cells were amassed and analyzed via flow cytometer (Becton Dickinson Co., Ltd., Maryland, USA), and the data were metered through MultiCycle for Windows (Beckman Coulter Life Sciences, Brea, CA, USA).
4
Cell Cycle Analysis of Treated Cells
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Cell cycle analysis was performed using a commercial kit (Coulter DNA Prep™ Reagents Kit; Beckman Coulter, Fullerton, CA, USA). Cells were plated in six-well plates and incubated overnight before treatment (Mock, 50 nmol/l dsCon and 50 nmol/l dsPAWR-435). Following treatment, cells were harvested, then washed twice with pre-chilled PBS and resuspended in 100 μl PBS at a concentration of 1×106 cells/ml. Each cell sample was mixed with 100 μl DNA Prep LPR (contained in Coulter DNA Prep™ Reagents Kit), gently mixed by vortex and incubated in the dark at room temperature (25°C) for 20 minutes. Then each was mixed with 1 ml of stain (DNA Prep Stain; contained in Coulter DNA Prep™ Reagents Kit), gently mixed by vortex and again incubated in the dark at room temperature (25°C) for 20 minutes. Finally, cell cycle analysis was performed within 1 hour using flow cytometry (Beckman Coulter FC500 Flow Cytometry System with CXP Software; Beckman Coulter, Fullerton, CA, USA), and the raw data was analyzed by Multicycle for Windows (Beckman Coulter).
5
Cell Cycle Analysis using Coulter DNA Prep
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Cell cycle analysis was performed using the Coulter DNA Prep™ Reagents Kit. Cells were plated in six-well plates and incubated overnight before treatment. After treatment for 24h, harvested cells were washed twice with prechilled PBS and resuspended with 100 μl PBS at a concentration of 1×106 cells/ml. Each cell sample was mixed with 100 μl DNA Prep LPR, gently mixed with a vortex, and incubated in the dark at room temperature (25°C) for 20 minutes. Then, each sample was mixed with 1 ml of DNA Prep Stain, mixed gently with a vortex, and incubated again in the dark at room temperature (25°C) for 20 minutes. Finally, the flow cytometry system (Beckman Coulter FC500 Flow Cytometry System with CXP Software; Beckman Coulter, Fullerton, CA, USA) was used for cell cycle analysis within 1 hour, and raw data was analyzed by Multicycle for Windows (Beckman Coulter).
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