Xtt assay protocol pdf files

Cell proliferation assays are widely used in cell biology for the study of growth factors, cytokines and media components, for the screening of. The xtt assay is used to measure cellular metabolic activity as an indicator of cell viability, proliferation and cytotoxicity. The use of xtt greatly simplifies the procedure of measuring proliferation, and is, therefore, an excellent solution to the quantitating of cells and their viability without using radioactive isotopes. The assay kit consists of two reagents, xtt reagent and electron coupling reagent.

Xtt labeling mixture to perform a cell proliferation assay xtt with one microplate 96 wells mix 5 ml xtt labeling reagent with 0. The cyquant xtt cell viability assay is a complete, optimized assay that generates a consistent colorimetric detection of viable mammalian cells. Xtt, commonly used in cell proliferation assays, is a tetrazolium compound. Prepare xtt working solution by combining xtt reagent with xtt activator according to the above.

Instructions for xtt reagent preparation and examples of applications. The optimal incubation time for this assay depends on experimental setup, such as. Xtt cell proliferation assay kit provides an easy to use tool for studying induction and inhibition of cell proliferation in any in vitro model. Mix each vial thoroughly to obtain a clear solution. Celltiter 96 aq ueous nonradioactive cell proliferation assay. Measurement of cell viability and proliferation comprise the underlying. Sample material is either adherent or suspension cells cultured in 96well microplates. Xtt assay for cell viability and proliferation sigma. Mtt proliferation assay protocol university of san diego. Cell proliferation kit ii xtt from roche biocompare. Please read these instructions carefully before beginning this assay. A collection of mtt assay protocols for research, provided by invitrogen. Xtt assay protocol continued 6 measure the absorbance of the assay wells containing the cells step 1a and the blank background. Xtt cell viability assay protocol cell signaling technology.

Add 50 l xtt detection solution to each well of 96well plate which contains 100200 lwell culture medium and return plate to incubator. Before the end of the treatment, remove the electron coupling reagent and the xtt reagent from the 20oc and thaw in a 37oc water bath. Xtt assay protocol for measuring cell viability, proliferation, activation and cytotoxicity. The cell proliferation kit ii xtt is a colorimetric assay for the nonradioactive quantification of cellular proliferation, viability, and cytotoxicity. Xtt is a negatively charged, tetrazolium salt that turns orange when it is reduced to a soluble. Thaw xtt labeling reagent and electroncoupling reagent, respectively in a waterbath at 37c. Xtt reagent is used to assess cell viability as a function of cellular redox potential. Remove cultures from incubator into laminar flow hood or other sterile working area. For most tumor cells, hybridomas, and fibroblast cell lines, 5,000 cells per well to perform proliferation assays. This kit was developed to assay cell proliferation in reaction to different growth factors. This colorimetric assay is based on the reduction of a yellow tetrazolium salt sodium 3. The atcc xtt cell proliferation assay kit incorporates a second generation.