Cell Proliferation Kit II (XTT)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- 1000 cells/ well were platted in a 96 well plate.	- Every 24h within a period of 96 h, cells were treated with XTT and incubated for 5 h.	- Absorbance values were measured at 490 nm

Upstream tips
- 1000 cells/ well were platted in a 96 well plate.
Protocol tips
- Every 24h within a period of 96 h, cells were treated with XTT and incubated for 5 h.
Downstream tips
- Absorbance values were measured at 490 nm

Publication protocol

The XTT Cell Proliferation Kit II (Roche Applied Science, Indianapolis, IN) was used following the manufacturer’s instructions. This assay is a nonwash colorimetric assay for cell proliferation and cell viability measurement. For this assay, an 2,3-Bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) tetrazolium salt is reduced by dehydrogenase enzymes in metabolically active cells in a soluble formazan, allowing direct measure of metabolic activity without removing the media from the plate. Briefly, 1000 cells of each cell line were plated onto 96-well plates in 100 μl of complete culture media. Every following 24 hours until 96 hours of growth, XTT reagent was added to each well, and the plates were incubated for 5 hours. Absorbance values were measured at 490 nm with a reference at 690 nm in a microplate reader (SpectraMax 190; Molecular Devices, Sunnyvale, CA). Experiments were performed in five replicates for each cell line at least five times (n = 5). Means were reported for the 24-hour absorbance value for each cell line.

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Manufacturer protocol

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