View Featured Offers >>
9095
XTT Cell Viability Kit
Cellular Assay Kits
Assay Kit
  1. Products
  2. Cellular Assay Kits
  3. XTT Cell Viability Kit

XTT Cell Viability Kit #9095

Citations (2)
Figure 1. C2C12 cells were seeded at varying density in a 96-well plate and incubated overnight. The XTT assay solution was added to the plate and cells were incubated. The absorbance at 450 nm was measured at 1.0, 2.0, 3.0, 4.0, and 5.0 hours.
Figure 2. C2C12 cells were seeded at 2x104 cells/well in a 96-well plate and incubated overnight. Cells were then treated with various concentrations of doxorubicin overnight. The cytotoxicity was measured using XTT Cell Viability Kit (red) and BrdU Cell Proliferation Assay Kit #6813 (blue) as shown in the left panel. The percentage inhibition in each assay was calculated and plotted in the right panel. Doxorubicin treatment can lead to cell DNA damage followed by cell cycle arrest.
Figure 3. HeLa cells were seeded at 1x104 cells/well in a 96-well plate and incubated overnight. Cells were then treated with various concentrations of staurosporine overnight. The cytotoxicity was measured using XTT Cell Viability Kit (red) and BrdU Cell Proliferation Assay Kit #6813 (blue) as shown in the left panel. The percentage inhibition in each assay was calculated and plotted in the right panel. Staurosporine is a nonspecific kinase inhibitor and induces cellular apoptosis.
Image Gallery
Learn more about how we get our images
To Purchase # 9095
Cat. # Size Qty. Price
9095S		 1 Kit  (1000 assays (96 well format))

Custom Formulation

Product Includes Quantity (with Count) Solution Color
XTT Reagent 2 x 25 ml
Electron Coupling Solution 2 x 0.5 ml Yellow

Protocol

PRINT

View >Collapse >

XTT Cell Viability Kit Protocol

A. Reagent Preparation

  1. Thaw reagents just before experiment.

NOTE: Precipitation may occur when reagents are stored at −20°C. Make sure reagents are clear prior to use. If necessary, warm reagents to 37°C to reconstitute.

  1. Add electron coupling solution to XTT Reagent (1:50 volume ratio) to make XTT detection solution. For example, each 96-well plate needs 5 ml XTT solution and 0.1 ml electron coupling solution.

B. XTT Assay

  1. Add 50 µl XTT detection solution to each well of 96-well plate (which contains 100–200 µl/well culture medium) and return plate to incubator.
  2. Read absorbance at 450 nm.

NOTE: The optimal incubation time for this assay depends on experimental setup, such as: cell type, cell number, and treatment. Optimization of incubation time can be determined by reading one plate at various time points after addition of XTT detection solution (featured below).

Figure 1. C2C12 cells were seeded at varying density in a 96-well plate and incubated overnight.

posted October 2012

Protocol Id: 46

Product Description

The XTT Cell Viability Assay Kit is a colorimetric assay that detects the cellular metabolic activities. During the assay, the yellow tetrazolium salt XTT is reduced to a highly colored formazan dye by dehydrogenase enzymes in metabolically active cells. This conversion only occurs in viable cells and thus, the amount of the formazan produced is proportional to viable cells in the sample. The formazan dye formed in the assay is soluble in aqueous solution and can be quantified by measuring the absorbance at wavelength 450 nm using a spectrophotometer. An electron coupling reagent, such as PMS (N-Methylphenazonium methyl sulfate), can significantly improve the efficiency of XTT reduction in cells.

Specificity / Sensitivity

The XTT Cell Viability Kit detects formazan dye produced from XTT conversion by mitochondrial enzymes in cells. Because these mitochondrial enzymes are inactivated shortly after cell death, the orange colored formazan dye only appears in viable cells. This XTT Cell Viability Kit is expected to work in most cells lines. Variable with cell type and incubation time employed in the assay, 0.2-2x104 cells/well should be sufficient for most experiments. For the best result, a cell number titration (as shown in Figure 1) is recommended.

Species Reactivity:

All Species Expected

Background

Cell viability and proliferation assays are widely used in drug discovery for the study of growth factors, cytokines, and cytotoxic agents. High throughput screening, in early drug discovery compound screening and in later drug safety and toxicity studies, requires a reliable, sensitive, and simple assay with the ability to analyze a large number of samples. Colorimetric cell viability assays using tetrazolium salt, such as MTT, XTT, WST-1, etc. were developed based on live cells reduction of tetrazolium salt into highly colored formazan compounds (1,2). In contrast to cell proliferation assays, such as radioactive thymidine or BrdU labeling of DNA in live cells followed by quantification of the incorporated thymidine (by quantifying sample radioactivity) or BrdU (using anti-BrdU antibody), the XTT assay doesn’t require radioactive materials, cell fixing, or cell permeabilization. Thus, unlike alternative cellular analysis assays, cells examined in the XTT assay may be used for further analysis.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not for Use in Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit our Trademark Information page.