CNE2 and CNE2-STC2-KO cells at approximately 60% confluence were exposed to X-radiation at doses of 0 and 4 Gy, respectively. Then, cells were digested with 0.25% trypsin in order to prepare single cell suspension. After washing with PBS, cells were stained with the cell cycle detection kit (BD Biosciences, San Diego, CA, USA) following the manufacturer’s instructions, and analyzed with an FACSCantoTMII flow Cytometer (BD Biosciences; San Jose, CA, USA). All analyses were repeated in triplicate.
Cell cycle detection kit
Manufactured by BD
Sourced in United States
The Cell Cycle Detection Kit is a laboratory tool designed to analyze and monitor the progression of cells through the different phases of the cell cycle. This kit provides the necessary reagents and protocols to quantify the distribution of cells in the G0/G1, S, and G2/M phases of the cell cycle.
Automatically generated - may contain errors
Lab products found in correlation
Facscalibur flow cytometer, by BD (5 mentions)
Facscalibur, by BD (3 mentions)
Novocyte flow cytometer, by Agilent Technologies (2 mentions)
Pt rnase staining solution, by BD (2 mentions)
70 μm falcon filters, by BD (2 mentions)
Cellquest software, by BD (2 mentions)
Facscanto 2 flow cytometer, by BD (1 mentions)
Lsrfortessa x 20, by BD (1 mentions)
Annexin 5 apoptosis detection kit, by BD (1 mentions)
Apoptosis detection kit, by BD (1 mentions)
Fitc annexin 5 apoptosis detection kit 1, by BD (1 mentions)
Facs cytometry, by BD (1 mentions)
Annexin 5 pe and 7 aad, by BD (1 mentions)
Multicaspase kit, by Merck Group (1 mentions)
Cell counting kit 8 assay, by Keygen Biotech (1 mentions)
Model 680, by Bio-Rad (1 mentions)
Modfit lt software, by BD (1 mentions)
Facscalibur system, by BD (1 mentions)
70 m cell strainer, by BD (1 mentions)
Sybr green pcr master mix, by Promega (1 mentions)
34 protocols using cell cycle detection kit
1
Cell Cycle Analysis of X-Irradiated Cells
2
Cell Cycle Analysis of Honokiol-Treated Pancreatic Cancer Cells
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PANC-1 or SW1990 cells were seeded onto the 6-well plates at a density of 27 cells/well. After being treated with 50 µM honokiol or vehicle for 24 h, cells were then fixed in 70% (v/v) cold ethanol at 4°C overnight. After washing, the fixed cells were collected and re-suspended in propidium iodide (PI)/RNase Staining Buffer (Cell Cycle Detection Kit, Signalway Antibody (SAB) Co. Ltd., Maryland, USA) and cell cycle was analyzed by using a flow cytometer (BD Biosciences, San Jose, USA).
3
Cell Apoptosis and Cycle Analysis
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Cell apoptosis and cell cycle distribution of the indicated cells were respectively determined using the cell cycle detection kit (BD Biosciences, San Jose, CA) or Annexin V Apoptosis Detection Kit (BD), according to the manufacturer’s instructions by a BD LSRFortessa X-20 (BD). The data were analyzed using FlowJo software version 10.4 (Tree Star, Inc., San Carlos, CA).
4
Apoptosis and Cell Cycle Analysis
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Cell Samples were harvested 24 hours after transfection of vectors or plasmids. Apoptosis detection kit (BD Biosciences, San Jose, CA, USA) was used to detect cell apoptosis by stained with fluorescein isothiocyanate (FITC)-Annexin V and propidium iodide (PI). For cell cycle analysis, samples were stained by PI alone using a cell cycle detection kit (BD Biosciences). Apoptosis and cell population rates at G0±G1, S, and G2±M phases were analyzed by flow cytometry.
5
Cell cycle and apoptosis analysis
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Cell seeding was done on six-well plate at 1 × 106 cells/well and grown to 70% confluence before treatment with TR35 at 0, 1, 2, and 4 mg/ml for 24 and 48 h. Cell cycle analysis was done using propidium iodide (PI) following manufacturer instructions (cell cycle detection kit, BD, 550825). Apoptosis analysis was done using a Annexin V-FITC Apoptosis Detection Kit I (BD Biosciences, 556547) and Annexin V-APC/PI Apoptosis Kit (Sungene Biotech, AO2001-11A-H) using manufacturer protocol and analyzed by flow cytometry on FACSCalibur (BD Biosciences). The flow cytometry data were analyzed using FlowJo X.
6
Cell Cycle Analysis by Flow Cytometry
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After treated with trypsin, the cells were collected and washed with PBS twice. Then, the cells were fixed with 70% cold ethanol at -20°C for 2 h, and passed through 70-μm Falcon Filters to obtain a mono-dispersed cell suspension. The mono-dispersed cells were incubated with RNase A and stained with propidium iodide (PI) at room temperature for 30 min (Cell Cycle Detection Kit, BD, USA). Flow-cytometric analysis was performed with a FACS Calibur flow cytometer (Becton Dickinson, Oxford, UK). Finally, the cell cycle would be analyzed utilizing Cell Quest Modfit software.
7
Cell Cycle and Apoptosis Analysis Protocol
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For cell cycle analysis, cells were seeded into 6-well plates, and then trypsinized and centrifuged. After washing in PBS, cells were treated with 70% of ice-cold ethanol for 2 h all night. The cell cycle detection kit (BD Bioscience, San Jose, CA, USA) was added for 30 min. For cell apoptosis analysis, cells in cold PBS were incubated with 100 μL of 1× binding buffer and 5 μL of Annexin V-PE and 7AAD (BD Biosciences) for 15 min in the dark. Both were analyzed by FACS cytometry (BD Biosciences).
8
Cell Cycle Synchronization and Analysis
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Cells were plated in six-well plates, followed by incubation with serum-free medium for 48 hours for cell cycle synchronization, and then recovered serum. Cells were collected at 2-time points, 0 and 24 hours, after serum recovery. The attached cells were harvested and washed with 1 ml PBS, then centrifuged at 1000 rpm for 10 min and discarded the supernatant. Fixed cells with 1 ml pre-cooled 75% ethanol, vortexed, and kept cells at 4 °C overnight. Then centrifuged cells at 1000 rpm for 10 min and discard the supernatant. Stained cells with cell cycle detection kit (BD Pharmingen, #554656) after washed with PBS, resuspend the cells in 0.5 ml PT/RNase staining solution (BD Pharmingen, #550825), 1 × 106 cells per tube, and incubated at room temperature in the dark for 15 min, then detect with NovoCyte Flow Cytometer (ACEA Biosciences).
9
Cell Cycle Analysis Protocol
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A Cell cycle detection kit was purchased from BD company. Adherent cells were digested with 0.25% trypsin and then collected into centrifuge tubes and washed three times with pre-cooled PBS to completely remove the trypsin. Cells were collected using a centrifuge at a 300g centrifugal force and 4 °C to avoid cell damage. The cells were resuspended by adding 500µl binding buffer and were then transferred to flow tubes. While vortexing (to loosen the pellet), 5 mL of cold 70 to 80% ethanol was added drop-by-drop into the cell pellet, and the cells were incubated at -20°C for a minimum of 2 hours. After the incubation, cells were washed twice to remove the ethanol. The washing was in 1X PBS. The cells were centrifuged for 10 minutes at 1,000 to 1,500 rpm, and then the supernatant was aspirated. The cells were incubated for 15 minutes at room temperature, and the cell cycle assay was performed with the Muse Cell Cycle assay kit, and the results were analyzed with flowjo_v10 software.
10
Apoptosis, Cell Cycle, and MTT Assay
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Apoptosis, cell cycle detection and MTT assay were performed as described previously 29 , 30 using the Multicaspase Kit (Millipore Corporation, Hayward, US), Cell Cycle Detection Kit (BD Pharmingen, San Diego, US) and Thiazolyl Blue Tetrazolium Bromide (MTT, Sigma‐Aldrich Corp.).
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