Detection of apoptosis was conducted using the Annexin V-FITC/PI apoptosis detection kit according to the manufacturer's protocol. Approximately 1–4 × 105 cells of LNCaP and DU-145 cells in 96-well plates at concentrations of 0, 20, 40 and 80 µM of 4-HNE were incubated in an incubator at 37°C with 5% CO2 for 24 h. Cells were digested by trypsin and harvested by trypsinization, washed in 1 × PBS and subsequently incubated for 15 min at room temperature in the dark in 500 µl of 1 × binding buffer containing 5 µl of Annexin V-FITC and 5 µl of propidium iodide (PI). Afterward, apoptosis was analyzed by a FACScan laser flow cytometer (FACSCalibur, Becton Dickinson, USA) and BD FACSDiva analysis software.
Facscan laser flow cytometer facscalibur
Manufactured by BD
Sourced in United States
The FACScan laser flow cytometer (FACSCalibur) is an instrument used for analyzing and sorting cells or particles in a fluid suspension. It utilizes laser technology to detect and measure various characteristics of cells, such as size, granularity, and fluorescence, as they pass through the instrument's flow chamber.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using facscan laser flow cytometer facscalibur
1
Apoptosis Detection in Prostate Cancer Cells
2
Cell Cycle Analysis of Breast Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The distribution of cell cycle was analyzed with PI staining by flow cytometry. MDA-MB-231 and MCF-7 cells were treated with/without TAIII (10 μM, 15 μM), TAIII (15 μM) + KU55933 (the inhibitor of p-ATM, cells were pretreated with 10 μM for 2 h), TAIII (15 μM) +SB203580 (the inhibitor of p-p38, cells were pretreated with 10 μM for 2 h). Following the treatment, cells were fixed by cold 75% ethanol at −20°C for overnight. The cells were then stained with PI/RNase buffer (BD Bioscience, San Diego, CA, United States). The cell cycle distribution was measured by FACScan laser flow cytometer (FACSCalibur, Becton Dickinson) or MACSQuant Analyzer 10 (Miltenyi Biotec, Bergisch Gladbach Germany), data analyzed with software MODFIT and CELLQUEST (Becton Dickinson, Franklin Lakes, NJ) or FlowJo_V10 (BD Bioscience, San Diego, CA, United States) software.
3
Cell Cycle Analysis of A549 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A549 cells were seeded in six-well plates by the density of 6 × 10 5 per well and kept overnight at 37°C for attachment. After treating with 0.1% DMSO or different concentrations of LXB-1 for 24 h, cells were harvested and washed twice with phosphate buffer solution (PBS), and fixed in 70% ethanol for 1 h. Fixed cells were washed with PBS before incubation with 0.5 mL PBS containing 0.05% RNase and 0.5% Triton X-100 for 30 min. The cells were then stained with 0.1 mg/mL propidium iodide (PI) and DNA content and cell cycle were determined using a FACScanlaser flow cytometer (FACSCalibur, Becton Dickinson, Franklin Lakes, NJ, USA). The data were analyzed using the software MODFIT and CELLQUEST (Vertion 2.2, BD Biosciences, Franklin Lakes, NJ, USA).
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!