This procedure was reported previously [32 (link)]. In brief, PC9 cells were cultured in 6-well culture plates at 3 × 105 cells/well and treated with increased doses of PPI for 24 h. Afterwards, the cells were washed and resuspended in PBS and incubated with 0.1 % sodium citrate containing propidium iodide (PI) 0.05 mg and 50 μg RNase for 1 h at room temperature (RT). The cells were washed and subjected to FACScalibur flow cytometric analysis (FC500, Beckman Coulter, FL, USA), and the proportion of cells within the G0/G1, S, and G2/M phases of the cell cycle were analyzed using the MultiCycle AV DNA Analysis software (Phoenix Flow Systems, Inc. San Diego, CA, USA).
Multicycle av dna analysis software
Manufactured by Phoenix Pharmaceuticals
Sourced in United States
MultiCycle AV DNA Analysis software is a laboratory equipment product that provides automated analysis of DNA samples. The software is designed to process and interpret DNA data, enabling researchers to efficiently analyze genetic information.
Automatically generated - may contain errors
Lab products found in correlation
Fc500, by Beckman Coulter (5 mentions)
Propidium iodide, by Thermo Fisher Scientific (3 mentions)
Rnase a, by Merck Group (2 mentions)
Cytomics fc500 flow cytometer, by Beckman Coulter (2 mentions)
Facs calibur flow, by Beckman Coulter (2 mentions)
Facscalibur, by Beckman Coulter (1 mentions)
Cytomics fc 500 mpl, by Beckman Coulter (1 mentions)
Rnase, by Merck Group (1 mentions)
C1052, by Beyotime (1 mentions)
16 protocols using multicycle av dna analysis software
1
Cell Cycle Analysis of PC9 Cells
2
Cell Cycle Distribution Analysis by PI Staining
3
Cell Cycle Analysis of NPC Cells
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This procedure was reported previously17 (link). Briefly, NPC cells were cultured in 6-well plastic plates at 2 × 105 cells/well and treated with increased doses of β-elemene for 24 h. Afterwards, the cells were harvested, washed with phosphate-buffered saline (PBS), and resuspended in 500 μL of cold PBS and ethanol (1.5 mL) for 2 h at 4 °C. Afterwards, the fixed cells were incubated in 1 mL of 0.1% sodium citrate containing propidium iodide (PI) and RNase for 30 min at room temperature. The cells were washed and subjected to FACS Calibur flow cytometric analysis (FC500, Beckman Coulter, FL, USA), and the proportion of cells within the G0/G1, S, and G2/M phases of the cell cycle were analyzed using the MultiCycle AV DNA Analysis software (Phoenix Flow Systems, Inc. San Diego, CA, USA).
4
Solamargine Induces NSCLC Cell Cycle Arrest
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This procedure was reported previously 12 , 36 . Briefly, NSCLC cells were cultured in 6‐well plates at 2 × 105 cells/well and treated with increased doses of solamargine for 24 hrs. Afterwards, the cells were harvested, washed and resuspended in cold PBS and ethanol for 2 hrs at 4°C. The fixed cells were incubated in 1 ml of 0.1% sodium citrate containing propidium iodide (PI) RNase for 30 min at room temperature. The cell cycle distribution was detected by flow cytometry (FC500; Beckman Coulter, FL, USA), and the percentage of cells within the G0/G1, S, and G2/M phases were analysed using the MultiCycle AV DNA Analysis software (Phoenix Flow Systems, Inc., San Diego, CA, USA).
5
Cell Cycle Distribution Analysis
6
Shikonin Induces Cell Cycle Alterations
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Hela and c33a cells were cultured in 6-well plates and treated with increased doses of shikonin for 24 h. Cell cycle experiment was carried out using the cell cycle staining kit based on the manufacture’s instruction. In brief, the cells were washed and resuspended in PBS and then incubated with 0.05 mg 0.1% sodium citrate containing propidium iodide and 50 µg RNase for 1 h at room temperature. The cells were washed and subjected to FACSCalibur flow cytometric analysis (FC500, Beckman Coulter, FL, USA). The proportion (percentage) of cells within the G0/G1, S, and G2/M phases of the cell cycle was analyzed using the MultiCycle AV DNA Analysis software (Phoenix Flow Systems, Inc. San Diego, CA, USA).
7
Cell Cycle Analysis by Flow Cytometry
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Transfected cells were trypsinized and counted 24 h post seeding. Cells were centrifuged at 900 xg and resuspended at 1 × 106 cells/ml in Dulbecco’s PBS. Cells were centrifuged again at 900 xg and fixed with chilled (− 20 °C) 70% ethanol with vortexing. Cells were then incubated at − 20 °C for at least 24 h then pelleted at 3000 xg. Cell pellets were washed twice with PBS then resuspended in 50 μl of 100 μg/ml RNase A (per 1 × 106 cells) and incubated at room temperature for 10 min. 400 μl of 50 μg/ml propidium iodide in PBS was added (per 1 × 106 cells) and incubated for 30 min. at 37 °C. DNA content, represented as propidium iodide fluorescence, was quantitated using a Cytomics FC 500MPL flow cytometer (Beckman Coulter) at excitation of 488 nm. FL1 (Em 525) was used to detect FITC fluorescence and FL3 (Em 610) was used to detect propidium iodide. FL3 histograms were analyzed using the MultiCycle AV DNA analysis software (Phoenix Flow Systems) available in the FCS Express 4 Plus-Research Edition program (De Novo Software). The histogram was fit with the SL G21 S0 one cycle fitting model. G2/G1 ratio was set at 1.93 and background was removed before G1, S and G2 DNA content was analyzed.
8
Cell Cycle Analysis by Flow Cytometry
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T47D and MCF-7 cells were quickly fixed with 70% methanol and incubated with a DNA-binding dye, propidium iodide (1 mM; Thermo Fisher Scientific), and 10 mg/mL RNAse (Sigma-Aldrich Co.) at 37°C for 30 minutes. DNA content was assessed with a two-laser BD Accuri C6 flow cytometer (Beckman Coulter, Miami, FL, USA). G0/G1, S, and M/G2 phases of the cell cycle distribution were then analyzed using a MultiCycle AV DNA Analysis software (Phoenix Flow Systems, San Diego, CA, USA).
9
Cell Cycle Analysis of HCC Cells
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This procedure was reported previously [31 (link)]. In brief, HCC cells were cultured in 6-well plates and treated with increased doses of UA for 24 h. Afterwards, the cells were harvested, and resuspended in 500 μL of cold PBS for 2 h at 4 °C. Following washes, the fixed cells were incubated in 1 mL of 0.1 % sodium citrate containing propidium iodide (PI) 0.05 mg and 50 μg RNase for 30 min at room temperature (RT), subjected to FACSCalibur flow cytometric analysis (FC500, Beckman Coulter, FL, USA). The proportion of cells within the G0/G1, S and G2/M phases was analyzed using the MultiCycle AV DNA Analysis software (Phoenix Flow Systems).
10
Cell Cycle Analysis of Liver Cancer Cells
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HCC cells were plated in 6-well plates at 2×105 cells per well and were subjected to 10 µM sorafenib, 20 nM bufalin and the combination of 10 µM sorafenib and 20 nM bufalin for 24 h at 37°C. Then cells were trypsinized by 0.25% trypsin and then washed with PBS. A cell cycle assay was applied (Beyotime Institute of Biotechnology; C1052). Subsequently, cells were fixed in 70% methanol at 4°C for 2 h and stained with propidium iodide (PI) [which consisted of 0.5 ml staining buffer, 25 µl PI staining reagent (20 X), 10 µl RNase A (50 X)] for 30 min at 37°C. A flow cytometer was applied (FC500, Beckman Counter, Inc., Brea, CA, USA) to detect fluorescence at excitation wavelength of 350 nm, The multiCycle AV DNA Analysis software (version 306; Phoenix Flow Systems, San Diego, CA, USA) was adopted to perform analysis.
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