PC-3 cells were plated in 96-well plates at a density of 3×103 per well until attached. Cells were transfected as described above, then treated with 100nM Triptolide or DMSO (negative control) for 24h. 10 μl cell counting kit-8 (CCK-8) solution (Beyotime, C0038), was added per well and the plate was incubated for 1 h at 37°C. The absorbance of each well was measured on an M200pro Multimode Plate Reader (Tecan) at 450 nm and 630 nm. Each treatment was performed in triplicate and experiments were repeated 3 times.
M200pro multimode plate reader
Manufactured by Tecan
Sourced in Switzerland
The M200pro Multimode Plate Reader is a versatile laboratory instrument designed for a wide range of absorbance, fluorescence, and luminescence measurements. It features a high-performance optical system and advanced software capabilities to provide accurate and reliable data for various applications.
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Lab products found in correlation
4 protocols using m200pro multimode plate reader
1
Triptolide Cytotoxicity in PC-3 Cells
2
Cell Viability Assay of Nitazoxanide and Tizoxanide
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The cell viability was assessed via an MTT assay as described in our previous studies17 (link), 18 (link), 19 (link). HepG2 cells were deposited in 96-well flat-bottom culture plates (5 × 103 cells/well), and exposed to various concentrations of nitazoxanide (Nit, 01088480, Adamas-beta, Shanghai, China) or tizoxanide (Tiz, HY-12687, MedChemExpress, USA) for 24 h. Media were then removed, 100 μL of 0.5 mg/mL MTT (M1025, Solarbio, Beijing, China) was added to each well. After 4 h incubation, the MTT reagent was discarded, and 180 μL of dimethyl sulfoxide (DMSO, BS087, Biosharp, Hefei, China) was added to dissolve the insoluble purple formazan product. The absorbance was then measured with Tecan Infinite M200PRO Multimode Plate Reader (Switzerland) at a wavelength of 490 nm.
3
Cytotoxicity Assay of Triptolide
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5000 cells per well were plated in 96-wells plate, cultured until attached, then treated with various doses of triptolide, using DMSO as the negative control and culture medium as the blank control. 24 h after treatment, 10 μl CCK-8 solution per well was added and the plate was incubated for 1 h at 37°C. The absorbance of each well was measured on an M200pro Multimode Plate Reader (Tecan) at 450 nm and 630 nm. Each treatment was performed in triplicate and experiments were repeated at least 3 times.
4
Triptolide-Induced Calcium Imaging in PC-3 Cells
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PC-3 cells were grown on sterilized glass coverslips overnight and treated with the indicated concentrations of triptolide for 24 h. The medium was then removed and the cells were washed with calcium-free wash buffer (135 mM NaCl, 2 mM KCl, 2 mM MgCl2, 10 mM HEPES, 4g/L glucose, pH 7.4). Equal volumes of 1 mM Fluo-4 AM and 20% pluronic F-127 were mixed and added to calcium-free wash buffer to prepare Fluo-4 AM loading solution. The final concentration of Fluo-4 AM was 5 μM. Cells were incubated in the Fluo-4 AM loading solution for 30 min at 37°C before the loading solution was removed. Cells were washed three time with calcium-free wash buffer, then incubated at 37°C for another 30 min. Data were analyzed with a BX51 + DP70 fluorescence microscope (Olympus) or an M200pro Multimode Plate Reader (Tecan).
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