Sie suchten nach: dimethylthiazol (Englisch - Italienisch)

the celltiter 96® aqueous one solution cell proliferation assay(a) is a colorimetric method for determining the number of viable cells in proliferation or cytotoxicity assays. the celltiter 96® aqueous one solution reagent contains a novel tetrazolium compound [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)- 2-(4-sulfophenyl)-2h-tetrazolium, inner salt; mts(a)] and an electron coupling reagent (phenazine ethosulfate; pes). pes has enhanced chemical stability, which allows it to be combined with mts to form a stable solution. this convenient “one solution” format is an improvement over the first version of the celltiter 96® aqueous assay, where phenazine methosulfate (pms) is used as the electron coupling reagent, and the pms solution and mts solution are supplied separately. the mts tetrazolium compound (owen’s reagent) is bioreduced by cells into a colored formazan product that is soluble in tissue culture medium (figure 1, 1). this conversion is presumably accomplished by nadph or nadh produced by dehydrogenase enzymes in metabolically active cells (2). assays are performed by adding a small amount of the celltiter 96® aqueous one solution reagent directly to culture wells, incubating for 1–4 hours and then recording the absorbance at 490nm with a 96-well plate reader the quantity of formazan product as measured by the absorbance at 490nm is directly proportional to the number of living cells in culture (figure 2). because the mts formazan product is soluble in tissue culture medium, the celltiter 96® aqueous one solution assay requires fewer steps than procedures that use tetrazolium compounds such as mtt or int (5,6). the formazan product of mtt reduction is a crystalline precipitate that requires an additional step in the procedure to dissolve the crystals before recording absorbance readings at 570nm (7). if you currently use a [3h]thymidine incorporation assay, addition of the celltiter 96® aqueous one solution reagent can be substituted for the pulse of [3h]thymidine at the time point in the assay when the pulse of radioactive thymidine is usually added. bioassay data comparing [3h]thymidine incorporation to the mts-based celltiter 96® aqueous assay and the original mtt-based celltiter 96® assay demonstrate that tetrazolium reagents can be substituted for [3h]thymidine incorporation figure 2. effect of cell number on absorbance at 490nm measured using the celltiter 96® aqueous one solution assay. various numbers of b9 hybridoma cells were added to the wells of a 96-well plate in rpmi containing 50μm 2-mercaptoethanol and supplemented with 5% fbs and 2ng/ml il-6. the medium was allowed to equilibrate for 1 hour, then 20μl/well of celltiter 96® aqueous one solution reagent was added. after 1 hour at 37°c in a humidified, 5% co2 atmosphere, the absorbance at 490nm was recorded using an elisa plate reader. each point represents the mean ± sd of 4 replicates. the correlation coefficient of the line was 0.993, indicating a linear response between cell number and absorbance at 490nm. the background absorbance shown at zero cells/well was not subtracted from these data.
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