To quantitatively assess the rate of apoptosis, AV-PI staining was conducted. We used Annexin V/propidium iodide double-labelled flow cytometry kit (KeyGen, Nanjing, China) as the manufacturer described. Briefly, 2 × 105 LX2 cells or L02 cells were treated with casticin (0–40 μM) for 12 h, respectively. After harvested and washed twice with PBS, the cells were resuspended in 500 μl binding buffer. Then, 5 μl of annexin V and 5 μl of PI were added. The mixture was incubated at room temperature for 5 min in the dark. Then cells were analysed by flow cytometry with a Becton Dickinson FACS-420 flow cytometer (Franklin Lakes, NJ, USA).
Facs420 flow cytometer
Manufactured by BD
Sourced in United States
The FACS420 is a flow cytometer that uses laser technology to analyze and sort cells or particles in a fluid sample. It is designed to measure various characteristics of individual cells, such as size, granularity, and fluorescence intensity, as the cells pass through the instrument's detection system.
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Lab products found in correlation
4 protocols using facs420 flow cytometer
1
Quantifying Apoptosis by AV-PI Flow Cytometry
2
Multiparametric Cell Characterization
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We used the following equipment: SK3c cell incubator (Sanyo Corporation, Japan); microplate reader (SpectraMaxM5, Molecular Device, USA); Gel Imaging System (UVP, USA), FACS420 Flow Cytometer (Becton-Dickinson, USA), Inverted fluorescence microscope (Olympus CKX71, Japan), and LB942 Microplate Luminometer (Berthold Technologics, Germany).
3
Assessing Cell Apoptosis Post-OGD
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At 0 h, 6 h, 12 h, 24 h, and 48 h after OGD induction, the cell apoptosis rates were assessed. The cells were treated with 0.5 ml trypsin (0.25%) for 30 min, and then re-suspended in 1 ml PBS. Next, they were incubated in 0.5 ml PI solution (10 μg/ml) at 37°C for 30 min in the dark. Apoptosis was determined using an FACS420 flow cytometer (Becton Dickinson, USA).
4
Quantifying Cellular Apoptosis via Flow
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After treatment for 24 h, the cells were digested and trypsinized, then harvested. The cells were treated according to the Annexin v-FITC/PI kit instructions (Biovision, USA). The cells were mixed with 400 μL binding buffer and then 4 μL PI and 4 μL Annesxin v were added. The cells were incubated at room temperature for 15–20 min at in dark room. FACS420 flow cytometer (Becton–Dickinson, USA) was used to detect cell apoptosis.
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