After TGF-β1 and/or HCPT treatment for 24 h, the fibroblasts were incubated with annexin V-fluorescein isothiocyanate (FITC) and propidium iodide (PI) working solution (BD Pharmingen, San Diego, CA, USA) for 30 min at room temperature. Analysis was performed using a flow cytometer (Beckman Coulter, Brea, CA, USA).
Pi working solution
Manufactured by BD
Sourced in United States
PI working solution is a laboratory reagent used in various biological and biochemical applications. It is a commonly used dye that binds to DNA, allowing for the visualization and quantification of nucleic acids. The core function of PI working solution is to provide a fluorescent labeling agent for the detection and analysis of DNA samples in laboratory settings.
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Lab products found in correlation
4 protocols using pi working solution
1
Annexin V-FITC and PI Apoptosis Assay
2
Apoptosis Assessment of HepG2 Cells
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After treated with HWE for 48 h, the HepG2 cells were fully digested by trypsin to obtain cell suspension. Cells were centrifuged at 4°C and 3,000 rpm for 10 min, then they were fully resuspended and washed. Then the cells were centrifuged at 4°C and 2,000 rpm for 10 min. We discarded the supernatant, added the binding buffer, annexin V-FITC, and PI working solution (BD pharmingen, United States) to each sample, mixed them evenly, and then filtered them with a 400 mesh filter to obtain single-cell suspension. The number of apoptotic cells was measured by flow cytometry.
3
Apoptosis Quantification in Endothelial Cells
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HUVECs were seeded at 2×105 cells/well in 24-well plates, incubated overnight, and treated with ox-LDL, Tim-3, anti-Tim-3, and/or selected signal transduction inhibitors. Cells were harvested and resuspended in 100 mL annexin-binding buffer containing 5 mL FITC-annexin V and 1 mL PI working solution (BD Biosciences, Franklin Lakes, NJ, USA), and then incubated in the dark for 15 minutes at room temperature. An additional 400 mL binding buffer was added, and HUVECs were analyzed immediately by flow cytometry (BD Biosciences).
4
Annexin V-FITC/PI Staining of DSCs
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Freshly isolated DSCs were seeded at 2 × 105 cells/well in 24-well plates; incubated overnight; and treated with TLR ligands, Tim-3, anti-Tim-3, and signal transduction inhibitors described above. The cells were harvested and resuspended in 100 μl annexin-binding buffer containing 5 μl FITC-annexin V and 1 μl PI working solution (BD Biosciences, Franklin Lakes, NJ, USA) and then incubated in the dark for 15 min at room temperature. An additional 400 μl binding buffer was added, and DSCs were analyzed immediately by flow cytometry (BD Biosciences).
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