Three ovarian cancer cell lines (A2780, A2780cis, and OVCAR-5), one breast cancer cell line (MDAMB-231), and one normal cell line (MRC-5) were grown in accordance with the supplier’s instructions and maintained at 37 °C in humidified atmosphere of 5% of CO2. In total, 1 × 103 cancer cells and 8 × 103 MRC-5 were seeded in a 96-well plate and treated after 24 h with six different concentrations of Pd(II) complexes (0.001, 0.01, 0.1, 1, 10, and 100 µM). Notably, stock solutions (10 mM) of all palladium complexes were prepared using DMSO as solvent. After 96 h from the treatment, cell viability was measured with a CellTiter-Glo assay (Promega, Madison, WI, USA) using Tecan M1000 or SynergyH1 microplate readers. IC50 values were calculated from logistical dose–response curves using GraphPad Prism 8 software. Averages were obtained from triplicates, and error bars are standard deviations.
Synergy h1
Manufactured by Tecan
Sourced in United States
The Synergy H1 is a multi-mode microplate reader designed for diverse applications in life science research. It features advanced optics and detection capabilities to provide reliable data across a wide range of assays. The core function of the Synergy H1 is to accurately measure absorbance, fluorescence, and luminescence signals from samples in microplates.
Automatically generated - may contain errors
Lab products found in correlation
Celltiter glo assay, by Promega (2 mentions)
Cck 8 solution, by Dojindo Laboratories (1 mentions)
Synergy h1, by Agilent Technologies (1 mentions)
Safire, by Tecan (1 mentions)
Clear viewseal sealer, by Greiner (1 mentions)
Lb 943 mithras2, by Tecan (1 mentions)
Il 12 il 23p40, by R&D Systems (1 mentions)
Hydrospeed, by Tecan (1 mentions)
Il 18, by Merck Group (1 mentions)
Tnf α, by R&D Systems (1 mentions)
6 protocols using synergy h1
1
Evaluating Anticancer Pd(II) Complexes
2
Growth Kinetics and Spot Assays
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Cells were diluted to approximately 0.01–0.02 OD660 (∼2-5 x105 cells) in 150μL YPD across 3–4 replicates. They were grown and read in a BioTek Synergy H1 or Sunrise Tecan microplate reader, with continuous double orbital shaking at 30°C or 37°C. Reads were taken while stationary every 10–20 min, and experiments were run for at least 24–36 h. For spot assays, strains were grown overnight in YPD at 30°C and spotted with serial dilutions on YPD agar. The plates were incubated for 2–3 days at 23°C, 30°C and 37°C.
3
CCK8 Assay for RGC Viability
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CCK8 solution (Dojindo Laboratories, Kumamoto, Japan) was added to 96-well plates (10 μl/well). After a 4-h incubation at 37°C, the plates were analyzed at 450 nm with the Tecan Genios microplate reader (Synergy H1, BioTAK). All values are expressed as the mean ± SD of at least three wells and at least three separate experiments. As the initial cell density was the same, higher OD values represent increased cell viability over time. We compared the changes in the OD values at different times to evaluate the RGC viability.
4
Cytotoxicity Evaluation of Pt(0) Complexes
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Two types of ovarian cancer cell lines (A2780 and A2780cis), one breast cancer cell line (MDA-MB-231) and a normal cell line (MRC-5) were cultured following the supplier’s guidelines (Sigma-Aldrich, St. Louis, MO, USA) and maintained at 37 °C in a humidified atmosphere containing 5% CO2. In 96-well plates, 1∙103 cancer cells and 8∙103 MRC-5 cells were seeded and treated after 24 h with six different concentrations of Pt(0) complexes (0.001, 0.01, 0.1, 1, 10, 100 µM) [30 (link),31 (link)]. Stock solutions (10 mM) of all platinum complexes were prepared using DMSO as a solvent. After 96 h of treatment, cell viability was assessed using a CellTiter-Glo assay (Promega, Madison, WI, USA) with Tecan M1000 or SynergyH1 microplate readers (Mennedorf, Switzerland). IC50 values were determined from logistical dose–response curves using GraphPad Prism 8 software. Triplicate measurements were taken to calculate averages, and standard deviations were represented by error bars.
5
Microplate-based Spectroscopic Analysis
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Measurements were carried out on several microplate readers (Tecan SAFIRE, Tecan, Switzerland; LB 943 Mithras2, Berthold, Germany; Synergy H1, BioTek, United States) at 25 °C. UV–vis spectra were recorded using a 96-well plate (transparent, polystyrene, flat bottom, Sarstedt, Germany) as sample holder with clear viewseal sealer (Greiner Bio-One, Austria). The spectra were obtained in the wavelength range from 340 nm to 500 nm or 800 nm for the hydrogels that release 4-nitrophenol or 5,5′dibromo-4,4′-dichloro-indigo, respectively. A 1 nm wavelength step size was using for the Tecan SAFIRE for enzyme detection and the LB 943 Mithras2 performing the E. coli DH5α strain detection, and a 2 nm was used for the Synergy H1 for the EHEC strain detection, respectively.
6
Cytokine Secretion Analysis by ELISA
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Cells were incubated for 24 h, pelleted, and supernatants were collected. Supernatants were analyzed for tumor necrosis factor alpha (TNF-α; R&D Systems; lower limit of detection—31.1 pg/mL), IL-12/IL-23p40 (R&D Systems; lower limit of detection—78.1 pg/mL), or IL-18 (Merck; lower limit of detection—81.92 pg/mL) by enzyme-linked immunosorbent assay (ELISA; plate reader: BioTek Synergy H1; Winooski, VT, USA; washer: Tecan HydroSpeed; Männedorf, ZH, Switzerland).
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