Cell viability was determined using MTT-based cell growth determination kit (Sigma) at 0, 24, 72, and 144 h. MTT solution was added to the cultures, which were then incubated at 37°C for 3 h. The culture medium was removed, and the formazan crystals were dissolved in isopropanol (MTT solvent). The plates were mixed for 10 min to enhance dissolution. Absorbance was measured at wavelength 550 nm with Victor 3 V multi-label reader (Perkin Elmer). Cell viability was represented as a percentage normalized with the uninfected cells at 0 h time point.
Victor 3 5 multi label reader
Manufactured by PerkinElmer
Sourced in United States
The Victor 3 V multi-label reader is a versatile and high-performance microplate reader that can measure a wide range of assays, including absorbance, fluorescence, and luminescence. It is capable of handling microplates with up to 384 wells and can perform rapid and accurate measurements. The Victor 3 V is designed to provide reliable and consistent results for researchers and laboratory professionals.
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Lab products found in correlation
5 protocols using victor 3 5 multi label reader
1
MTT-based Cell Viability Assay
2
Cell Proliferation Assay Protocol
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MTT and PrestoBlue assays were used to determine proliferation rate. For both assays 3000 cells/well were seeded in 96-well plates in a final volume of 100 μL/well. For the MTT assay, the growth medium was replaced by growth medium supplemented with 0.5 mg/mL 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma M5655) for 3 hours at 37 °C, 16 (t1) and 40 h (t2) after seeding. The absorbance was measured at 595 nm, as described previously [43 (link)]. For the PrestoBlue assay, the growth medium was replaced with growth medium supplemented with 4% PrestoBlue Cell Viability Reagent (ThermoFisher, A13261). After 3 h of incubation at 37 °C, fluorescence was measured at 590 nm as a read out for mitochondrial respiration on a victor3V multilabel reader (Perkin Elmer, Waltham, MA, USA). Three technical replicates were included in each experiment and assays were repeated at least twice.
3
Nitric Oxide Quantification Protocol
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Nitric oxide (NO) concentration in supernatant was assessed using a Nitrate/Nitrite Colorimetric Assay Kit (Cedarlane, Missisauga, ON, Canada). The absorbance was measured at 540 nm with a Victor 3V multilabel reader (PerkinElmer, Waltham, MA, United States).
4
Fibrinogen Coagulation Assay with DHLA
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Fibrinogen coagulation assay was performed with fibrinogen at the final concentration of 0.34 μM, alone or in the presence of DHLA at final concentrations of 0.34 μM or 0.68 μM. Reaction mixtures also contained CaCl 2 at the final concentration of 2.2 mM and human thrombin, 1 IU/mL. Coagulation assay was carried out at room temperature in a microtiter plate. Immediately after the addition of thrombin, the microplate was placed in Victor 3 V multilabel reader (PerkinElmer, USA) and the rise in the absorbance at 350 nm was continually followed for 8 min. The absorbances of reaction mixtures without fibrinogen were also recorded and used to correct the corresponding results obtained in the presence of fibrinogen. The results were analysed according to published procedure [22] (link).
Journal Pre-proof Prepared protein and ligand were the starting point for Induced Fit Docking protocol [23, (link)24] (link).
Potential binding sites were determined using Maestro Sitemap [25] module and standard Induced Fit Docking protocol was used to produce up to 20 protein-ligand complexes. The obtained complexes were evaluated and the best docking orientation was selected based on the number of protein-ligand interactions and the docking score. Results were visualised using Schrodinger Maestro software.
Journal Pre-proof Prepared protein and ligand were the starting point for Induced Fit Docking protocol [23, (link)24] (link).
Potential binding sites were determined using Maestro Sitemap [25] module and standard Induced Fit Docking protocol was used to produce up to 20 protein-ligand complexes. The obtained complexes were evaluated and the best docking orientation was selected based on the number of protein-ligand interactions and the docking score. Results were visualised using Schrodinger Maestro software.
5
Evaluating TiO2 NP Cytotoxicity via Alamar Blue
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The Alamar Blue reagent, a water-soluble resazurin dye, was used to evaluate cell viability as a function of resazurin reduction to red fluorescent dye resofurin by metabolically active cells. BEAS-2B cells were seeded in transparent 96-well plates and exposed to NM100, NM101 or NM103 TiO2 dispersions at mass concentrations ranging from 2 to 100 µg/ml (0.62–31.25 µg/cm2) for 3, 24 or 48h. After exposure, cells were washed and a 10% Alamar Blue solution (ThermoFisher Scientific) was added in each well and incubated for 2h at 37°C. Blank control wells without cells were prepared for each particle dispersion to exclude possible interactions with the assay. In these control wells, the Alamar Blue reagent was directly added to reach a 10% final concentration. Cells treated with 0.1% Triton X-100 (Sigma–Aldrich) was used as a positive control. The fluorescence was measured on a VICTOR3 V multilabel reader (PerkinElmer) with an excitation wavelength of 560nm and an emission wavelength of 590nm. The fluorescence values were normalised by the controls and expressed as percent viability.
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