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a Shows percent change in the metabolic activity of S. mutans using MTT assay with various concentrations of test compounds.
The PI penetration in Candida cells treated with various concentrations of test compounds and positive control is shown in Figure 6.
Five increasing concentrations of test compounds were assayed to obtain % inhibition of the enzymatic activity in the range of 20 80.
IC50 values were calculated by measuring the effect of varying concentrations of test compounds by using the EZ-Fit Enzyme Kinetic program.
Estimation of IC 50 values: The concentrations of test compounds which inhibited the hydrolysis of substrates (acetylthiocholine and butyrylthiocholine) by 50% (IC 50 ) were determined by monitoring the effect of increasing concentrations of these compounds in the assays on the inhibition values.
The mutant protein TTRY78F was incubated with various concentrations of test compounds in 96 well microplates.
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Assay mixture (180 μL) containing three different concentrations of test compound was added to three rows of a microtitre plate, a fourth row contained only the assay mixture.
In brief, different concentrations of test compound were prepared by serial dilution and 20 μl of each concentration was added to the rows of a 96-well microtitre plate in diminishing concentrations.
Logarithmically growing cells were plated onto 96-well plates (Falcon, Marathon Lab Supplies, London, UK) at a density of 5 6 × 10 cells per well four hours before treatment with a range of concentrations of test compound in tetrahydrofuran (THF; Sigma) vehicle.
Aliquots of the assay mixture (180 μL) containing three different concentrations of test compound were added to three rows of a microtitre plate, a fourth row contained only the assay mixture.
The bone resorption assay is a functional in vitro assay that measures Type-I collagen degradation after a three-day incubation of rabbit osteoclasts cultured on bovine bone with varying concentrations of test compound, as previously described [ 23].
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