For cell cycle analysis, ~2×106 cells were harvested and fixed overnight with 70% ethanol at −20°C. After washing twice with PBS, the cells were suspended in clean PBS with 50 μg/ml propidium iodide and 10 μg/ml RNase A for 30 min in the dark at room temperature. The cells were then analyzed using FACStar Flow Cytometry (BD Biosciences), and the cell cycle distribution was analyzed.
Facstar flow cytometry
Manufactured by BD
The FACStar is a flow cytometry instrument manufactured by BD. It is designed to analyze and sort cells or other particles in a fluid sample. The FACStar utilizes laser technology to detect and measure various physical and fluorescent characteristics of individual cells or particles as they pass through the instrument's flow cell.
Automatically generated - may contain errors
3 protocols using facstar flow cytometry
1
Cell Cycle Analysis by Flow Cytometry
2
Propidium Iodide Cell Cycle Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell cycle was analysed by propidium iodide (Nanjing, China) (PI) staining. Briefly, transfected gastric cancer cells were cultured in a 6-well plate at a density of 10 5 cells/well and cultured for 24 hours. After fixation with 70% ethanol at 4°C for 2 hours and PI staining, cells were analysed by FACStar flow cytometry (BD Biosciences).
3
Measurement of Mitochondrial Membrane Potential
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ΔѰ m was measured using the Rhodamine 123 mitochondrial-specific fluorescent dye (Sigma-Aldrich; Merck KGaA; Ex/Em=485/535 nm), as previously described (23) . Briefly, 1x10 6 cells in 60 mm culture dish (Nalge Nunc International) were incubated with the indicated amounts of GA with or without Z-VAD-FMK, NAC or BSO at 37˚C for 24 h. Cells were washed twice with PBS and incubated with Rhodamine 123 (0.1 µg/ml) at 37˚C for 30 min. Rhodamine 123 staining intensity was determined by FACStar flow cytometry (BD Biosciences) and analyzed using CellQuest Pro software (version 5.1; BD Biosciences). An absence of Rhodamine 123 from cells indicated the loss of ΔѰ m in CPAECs.
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!