ROCHE Cell Proliferation Kit II (XTT)

Stock Code: 2795617
Manufacturer Part No: 11465015001

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Descriptions

Application

The Cell Proliferation Kit II (XTT) is used for the nonradioactive, spectrophotometric quantification of cell proliferation and viability in cell populations using the 96-well-plate format. It can be used for: Measurement of cell proliferation in response to growth factors, cytokines, mitogens, and nutrients.Analysis of cytotoxic and cytostatic compounds, such as anti-cancer drugs and other pharmaceutical compounds.Assessment of growth-inhibitory antibodies and physiological mediators that inhibit cell growth.Testing of biocompatibility of various scaffolds, employed in bone tissue engineering, for bone cell growth.cell viability assay.

Features and Benefits

Safe and easy: Eliminate radioactive isotopes, washing steps, and additional reagents. Accurate: The absorbance obtained strongly correlates to the cell number. Sensitive: Detect low cell numbers. Fast: Process a large number of samples using a multi-well ELISA reader.

General description

The Cell Proliferation Kit II (XTT) is a colorimetric assay for the nonradioactive quantification of cellular proliferation, viability, and cytotoxicity. Sample material is either adherent or suspension cells cultured in 96-well microplates.Colorimetric assays analyze the number of viable cells by the cleavage of tetrazolium salts added to the culture medium. This technique requires neither washing nor harvesting of cells, and the complete assay, from microculture to data analysis by an ELISA reader, is performed in the same microplate.More recently, the tetrazolium salt XTT was described. In contrast to MTT, the cleavage product of XTT is soluble in water; therefore, a solubilization step is not required. The tetrazolium salt XTT is cleaved to formazan by a complex cellular mechanism. This bioreduction occurs in viable cells only, and is related to NAD(P)H production through glycolysis. Therefore, the amount of formazan dye formed directly correlates to the number of metabolically active cells in the culture.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Packaging

1 kit containing 2 components.

Preparation Note

Working solution: Preparation of solutionsThaw XTT labeling reagent and electron-coupling reagent, respectively in a water bath at 37 °C. Mix each vial thoroughly to obtain a clear solution.XTT labeling mixtureTo perform a cell proliferation assay (XTT) with one microplate (96 wells) mix 5 ml XTT labeling reagent with 0.1 ml electron coupling reagent.Note: To obtain reliable results thaw and mix XTT labeling reagent and electron coupling reagent immediately before use.Working instructionCells are grown in microplates (tissue culture grade, 96 wells, flat bottom) in a final volume of 100 µl culture medium per well, according to the media needs of the cells in a humidified atmosphere (e.g., 37 °C, 6.5% CO₂).The incubation period of the cell cultures depends on the particular experimental approach and on the cell line, used for the assay. For most experimental setups, the incubation of cells for 24 to 96 hours is appropriate