Orangu™ Cell Counting Solution
Orangu™ is a non-cytotoxic, highly sensitive, colorimetric assay for the determination of cell viability in cell proliferation and cytotoxicity assays. Orangu™ utilizes WST-8 to give an orange-coloured formazan dye. The amount of dye formed is directly proportional to the number of living cells. Increasing incubation periods or number of cells will lead to an increase in the amount of orange dye. Simple!
The assay utilizes WST-8, a highly water-soluble tetrazolium salt, which produces an orange coloured formazan dye upon reduction in the presence of an electron mediator. The amount of the formazan dye generated by dehydrogenases in the cells is directly proportional to the number of viable cells and the length of incubation.
Orangu™ relies on the activity of multiple intracellular dehydrogenases while other proliferation quantification methods rely only on the activity of the mitochondrial dehydrogenase.
Chemical reaction from WST-8 to WST-8 formazan.
The Orangu™ assay is supplied as a ready-to-use solution and can be performed in 3 steps:
Step 1. Add neat Orangu™ solution to each well
Step 2. Incubate the plate for 1 to 4 hours in a humidified incubator
Step 3. Measure the absorbance at 450 nm using a standard microplate reader
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Orangu™ demonstrates higher levels of sensitivity than commonly used alternatives MTT, XTT, MTS or WST-1, guaranteeing that even small cell number differences are detected.
Cell proliferation assay comparing Orangu™ (blue), MTS (pink), XTT (green), and MTT (grey). Increasing numbers of HeLa cells were incubated in MEM media containing FCS and the different proliferation reagents for 2 hours. Compared to the other cell counting reagents, Orangu™ is linear over a broader range, demonstrating accurate cell quantification.
Cell viability study on HeLa cells (5000 cells/well) comparing the toxicity of Orangu™, WST-1 and MTS assays at different time points (1, 12 and 24 hours). Orangu™ is the only non-toxic assay as viable cells are clearly visible up to 24 hours. In contrast, in both the WST-1 and MTS assay, only round non-viable cells are observed at the same time point.