To examine the cell cycle distribution of asynchronous populations of lung cancer cells, replicative DNA synthesis and DNA content were analyzed using bivariate flow cytometric analysis. Parkin siRNA- or mutant parkin (R275W or G430D)-transfected A549 cells were harvested by trypsin-EDTA release and fixed in ice-cold 70% ethanol. At least 1 to 2 h before flow cytometric analysis, cells were resuspended in a 1 mL aliquot of modified Vindelov's DNA staining solution (10 μg/mL RNase A and 5 μg/mL propidium iodide in PBS). Flow cytometric analysis was performed with the flow cytometry system (FACS Calibur-S System; BD Biosciences, San Jose, CA). Cells in the G1, S, and G2-M phases of the cell cycle were determined with Modfit LT (Verity House Software, Top-sham, ME).
Facs calibur s system
Manufactured by BD
Sourced in Germany
The FACS Calibur-S System is a flow cytometry instrument designed for cell analysis and sorting. It features a compact design and offers multicolor detection capabilities. The system provides precise and reliable data acquisition for a variety of applications in life science research and clinical settings.
Automatically generated - may contain errors
Lab products found in correlation
8 protocols using facs calibur s system
1
Cell Cycle Analysis of Lung Cancer Cells
2
Quantifying Apoptosis by Annexin V/PI Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptotic cells were quantified by Annexin V/PI double staining assay using apoptotic detection kit (BioVision). H1299 cells were seeded in 6‐well plates at a density of 1 × 105 cells/well. After overnight incubation, cells were treated with SFE at 0 or 500 μg/ml in serum‐complete media for 72 hr. Cells were then gently detached by brief trypsinization, washed, and stained with annexin V‐FITC and PI for 10 min in the dark. The stained cells were analyzed using flow cytometer (FACS Calibur‐S System, BD Biosciences). Annexin V‐positive/PI‐negative cells and annexin V‐positive/PI‐positive cells were identified as early and late apoptotic cells, respectively.
3
Cell cycle analysis by flow cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RWPE-1 cells were harvested by trypsin-EDTA and fixed in ethanol (70% in PBS). At least 1 to 2 h before flow cytometry, cells were resuspended and then stained with 10 μg/mL RNase A and 5 μg/mL propidium iodide in PBS. Cell distribution in the different phases of the cell cycle was analyzed by flow cytometry (FACSCalibur-S System, BD Bioscience) equipped with FlowJo software.
4
Ebractenoid F-Induced Cell Cycle Arrest in Lung Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To examine the cell cycle arrest of lung cancer cells, ebractenoid F-treated A549 and H460 cells were seeded on a 12-well plate, harvested by trypsin, pelleted, re-suspended in PBS, and fixed in 70% ethanol. At least 1–2 h before the flow cytometric analysis, the cells were washed in PBS and stained with 500 μL of PI/RNase (Propidium Iodide) solution. The flow cytometric analysis was performed with the flow cytometry system (FACS Calibur-S System; BD Biosciences, San Jose, CA, USA).
5
Phenotypic Characterization of TMSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
TMSCs were phenotypically characterized by flow cytometry. The TMSCs (1.0 × 104 cells) from the two experimental groups were incubated with fluorescein isothiocyanate (FITC)- or phycoerythrin (PE)-conjugated monoclonal antibodies against Isotype-PE, Isotype-FITC, CD14, CD34, CD45, CD73, CD90, and CD105 (BD Biosciences, San Jose, CA, USA) for 30 min at 4 °C. The cell populations were analyzed using a FACScan instrument (FACSCalibur-S System; BD Biosciences). A total of about 1 × 104 cells were counted, of which 9832 were live cells except of dead cell and debris. As a control, non-treatment TMSCs and isotype-PE and isotype-FITC Ig control for each wavelength were used. Data were analyzed using Flowjo (BD Biosciences). Results were displayed as the percentage of cells labeled for each monoclonal antibody.
6
MLE Modulates Cell Cycle Progression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To determine the effect of MLE on cell cycle, starved HCT116 and H1299 cells (3 × 105 cells/well) were cultured in serum complete media containing 0 or 700 µg/ml of MLE for 72 h (HCT116) or 48 h (H1299). Cells were then harvested by trypsinization, fixed in 70 % ethanol overnight at -20 °C, incubated with Ribonuclease A (1 µg/ml; Sigma-Aldrich) and propidium iodine (50 µg/ml; Sigma-Aldrich) for 20 min in the dark at room temperature. Cell population at Sub-G1, G1/G0, S, and G2/M phases were identified according to the DNA content using a flow cytometer (FACS Calibur-S System, BD Biosciences, Heidelberg, Germany) and Cell Quest Pro software (BD Biosciences).
7
Cell Cycle Analysis in Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell cycle was determined as previously described [25 (link)] with a slight modification. Briefly, HCT116 and H1299 cells (3 × 105 cells/well) were seeded in 10-cm culture dishes (Corning Inc.). After 24 h, cells were synchronized for another 24 h in serum free media. Cells were then treated with PLE at 350 µg/ml in serum complete media for 24 h (in the case of H1299) or 72 h (in the case of HCT116). Adherent cells were detached by brief trypsinization (0.25% trypsin-EDTA; Sigma-Aldrich). Cell pellets were washed with ice-cold PBS and then re-suspended in 70% ethanol overnight for fixation. After centrifugation at 3,000 × g, the supernatant was removed, and cells were incubated with PBS containing 1 µg/ml RNAse (Sigma-Aldrich) and 50 µg/ml propidium iodine (Sigma-Aldrich) for 20 min at room temperature. Single-cell suspension was generated by gentle pipetting. Cell cycle analysis was performed using a flow cytometer (FACS Calibur-S System, BD Biosciences, Heidelberg, Germany), and data were processed using Cell Quest Pro software (BD Biosciences).
8
Cell Cycle Analysis of Mesenchymal Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cell cycle stage was examined by measuring the DNA content of nuclei labeled with propidium iodide (PI, Abcam). For the cell cycle analysis, either young or old TMSCs (1 × 105 cells) were harvested by trypsinization and gently pelleted by centrifugation at 300 × g for 5 min. The pellets were washed and resuspended in 200 μl DPBS. The resuspended cells were transferred dropwise into 70% ethanol (800 μl) and fixed for 1 week. The fixed cells were collected, washed, and resuspended in PI staining solution (50 mg/ml) containing RNase A (100 mg/ml) and incubated in the dark for 30 min at room temperature. A FACScan (FACSCalibur-S System; BD Biosciences) was used to analyze the cell cycle, and the FlowJo software was used for data analysis.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!