2013 Promega catalogue

Life
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2013
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Cell Signaling
CellTiter 96 ® AQ ueous Non-Radioactive Cell
Proliferation Assay (MTS)
Product Size Cat.# Price ($)
CellTiter 96 ® AQ ueous Non-Radioactive Cell
Proliferation Assay
1,000 assays G5421 394
5,000 assays G5430 1124
50,000 assays G5440 Pls. Enq.
Available Separately Size Cat.# Price ($)
CellTiter 96 ® AQ ueous MTS Reagent Powder 1 g G1111 1364
For Laboratory Use.
250 mg G1112 424
Description: The CellTiter 96 ® AQ ueous Non-Radioactive Cell Proliferation
Assay is a homogeneous, colorimetric method for determining the number
of viable cells in proliferation, cytotoxicity or chemosensitivity assays. The
CellTiter 96 ® AQ ueous Assay is composed of solutions of a novel tetrazolium
compound [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-
sulfophenyl)-2H-tetrazolium, inner salt; MTS] and an electron coupling reagent
(phenazine methosulfate) PMS. MTS is bioreduced by cells into a formazan
product that is soluble in tissue culture medium. The absorbance of the
formazan product at 490nm can be measured directly from 96-well assay
plates without additional processing. The conversion of MTS into the aqueous
soluble formazan product is accomplished by dehydrogenase enzymes found in
metabolically active cells. The quantity of formazan product as measured by the
amount of 490nm absorbance is directly proportional to the number of living
cells in culture.
If you currently use a [ 3 H]-thymidine incorporation assay, addition of the
combined MTS/PMS solution can be substituted for [ 3 H]-thymidine at the time
point in the assay when the pulse of radioactive thymidine is usually added.
Data from proliferation bioassays comparing the CellTiter 96 ® AQ ueous Assay
and [ 3 H]-thymidine incorporation show similar results.
CellTiter 96 ® AQ ueous MTS Reagent Powder is a novel tetrazolium
compound for use in colorimetric assays for determining the number of viable
cells in proliferation, cytotoxicity or chemosensitivity assays. It is provided in
powdered form.
Features:
• Easy to Use: Combine provided MTS and PMS solutions, add to cells,
incubate and read absorbance.
• Fast: Perform the assay in a 96-well plate with no washing or cell harvesting.
Also eliminates solubilization steps because the MTS formazan product
is soluble in tissue culture medium.
• Non-Radioactive: Requires no scintillation cocktail or radioactive waste
disposal (unlike [ 3 H]-thymidine).
• Flexible: Plates can be read and returned to incubator for further color
development (unlike MTT).
• Safe: Requires no volatile organic solvent to solubilize the formazan
product (unlike MTT).
Storage Conditions: For long-term storage, store MTS and PMS Solutions at
–20°C, protected from light.
CellTiter 96 ® Non-Radioactive Cell
Proliferation Assay (MTT)
Product Size Cat.# Price ($)
CellTiter 96 ® Non-Radioactive Cell Proliferation 1,000 assays G4000 371
Assay
5,000 assays G4100 1085
For Laboratory Use.
Description: The CellTiter 96 ® Assay is a collection of qualified reagents that
provide a convenient method of determining viable cell number. The CellTiter
96 ® Assay is a modification of the MTT assay method described by Mosmann
and incorporates several improvements to the method that address previous
technical problems including: 1) serum protein precipitation caused by adding
organic solvent; 2) interference by phenol red; 3) incomplete solubilization
of the formazan crystals resulting in lower sensitivity; and 4) stability of the
colored product.
Perform the CellTiter 96 ® Assay by adding a premixed, optimized Dye Solution
to culture wells of a 96-well plate, usually containing various concentrations
of growth factor or test substance. During a 4-hour incubation, living cells
convert the MTT tetrazolium component of the Dye Solution into a formazan
product. If you currently use a [ 3 H]-thymidine incorporation assay, the addition
of Dye Solution can be substituted for the pulse of radioactive thymidine at the
time point in the assay when the pulse of [ 3 H]-thymidine is usually added. The
Solubilization/Stop Solution is then added to the culture wells to solubilize the
formazan product, and the absorbance at 570nm is recorded using a 96-well
plate reader. Direct comparison between [ 3 H]-thymidine incorporation and
tetrazolium conversion ha demonstrated less than a 5% difference between the
two assays for determination of growth factor content of several samples.
Features:
• Gain Sensitivity: Detect as few as 1,000 cells/well with a 96-well plate
reader. Greater sensitivity than the neutral red assay procedure.
• Use a Variety of Cells: Assay mammalian, plant and yeast cells.
• Non-Radioactive: Requires no scintillation cocktail or radioactive waste
disposal.
• Save Time: Perform the assay in a 96-well plate with no washing steps,
no cell harvesting and no scintillation counting.
• Adapt to Your Needs: Follow either a 4-hour or overnight protocol.
• Convenient: Requires no weighing or mixing of dye components.
Storage Conditions: Store Dye Solution at –20°C and Solubilization/Stop
Solution at room temperature.
Protocol
CellTiter 96 ® Non-Radioactive Cell Proliferation Assay Technical
Bulletin
Part#
TB112
Protocol
CellTiter 96 ® AQ ueous Non-Radioactive Cell Proliferation Assay
Technical Bulletin
Part#
TB169
Section
Contents
Table of
Contents
48
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