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CT02 MTT Cell Growth Assay Kit

CT02
1000 assays  
Purchase on Sigma-Aldrich

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Key Applications
ACT
Description
Catalogue NumberCT02
Brand Family Chemicon®
Trade Name
  • Chemicon
DescriptionMTT Cell Growth Assay Kit
OverviewMTT is a pale yellow substrate that is cleaved by living cells to yield a dark blue formazan product. This process requires active mitochondria, and even freshly dead cells do not cleave significant amounts of MTT. The colorimetric assay described below can be used for either proliferation or complement-mediated cytotoxicity assays.
Materials Required but Not DeliveredIsopropanol with 0.04 N HCl for color development
References
Product Information
Components
  • Reagent A: MTT, (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide), 50 mg/vial.
  • Solution B: PBS pH 7.4, 15 mL
HS Code3822 19 90
Quality LevelMQ100
Applications
ApplicationMTT Cell Growth Assay is a colorimetric assay that can be used for either proliferation or complement-mediated cytotoxicity assays.
Key Applications
  • Activity Assay
Application NotesProcedure:
1) Carry out a lymphokine, mitogen, or complement-mediated cytotoxicity assay using standard methods, in 96-well flat-bottomed tissue culture plates of good optical quality (e.g. Falcon). The final volume of tissue culture medium in each well should be 0.1 mL, and the medium (e.g. RPMI or DMEM) may contain up to 10% Fetal Bovine Serum.

2) At the end of the assay add 0.01 mL AB Solution (MTT) to each well. Mix by tapping gently on the side of the tray.

3) Incubate at 37° C for cleavage of MTT to occur. Optimal times may vary according to the assay, but four hours is suitable for most purposes. At the end of this time, the MTT formazan produced in wells containing live cells will appear as black, fuzzy crystals on the bottom of the well.

4) Add 0.1 mL isopropanol with 0.04 N HCl to each well. Mix thoroughly by repeated pipetting with a multichannel pipettor. The HCl converts the phenol red in tissue culture medium to a yellow color that does not interfere with MTT formazan measurement. The isopropanol dissolves the formazan to give a homogeneous blue solution suitable for absorbance measurement.

5) Within an hour, measure the absorbance on an ELISA plate reader with a test wavelength of 570 nm and a reference wavelength of 630 nm. After several hours at room temperature, serum proteins may begin to precipitate due to the acid/alcohol. Chilling the plates will hasten the precipitation. If the plates must be stored before measuring, keep at 4° C before adding the acid/alcohol, then warm to room temperature and add acid/alcohol just before reading.

Results:
The MTT assay will normally detect 200 to 50,000 cells of a typical cell line, although 1,000 to 50,000 is the useful range. This number may vary for other cell types. Cytotoxic assays should be set up so that the control, unlysed cells give a signal of 0.2 to 0.4, and proliferation assays should yield a similar value at plateau concentrations. This corresponds to about 20-50,000 cells per well with a typical cell line.

Absorbance is directly proportional to the number of cells; actual cells do not absorb significantly, even up to concentrations of 1 x 106 cells/mL.
Biological Information
Species Reactivity
  • Vertebrates
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Usage Statement
  • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage ConditionsMaintain at 2-8°C for up to six months. The Reagent A / Solution B mixture is stable at 2-8°C for up to two weeks.

Reagent Preparation:
Add 10 mL Solution B to one vial of Reagent A. Mix well, sterile filter and keep in the dark at 4° C until used. Note: It may take overnight to dissolve. Do not heat solution. When absolutely required to dissolve crystals, adjust pH with 1-2 drops of HCl. The AB mixture is stable for several weeks under these conditions.
Packaging Information
Material Size1000 assays
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Número de referencia GTIN
CT02 08436037123979

Documentation

MTT Cell Growth Assay Kit Ficha datos de seguridad (MSDS)

Título

Ficha técnica de seguridad del material (MSDS) 

MTT Cell Growth Assay Kit Certificados de análisis

CargoNúmero de lote
COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION 2952750
COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION 3138395
COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION 3076703
COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION 3100161
COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION 3023724
COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION 3060580
COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION 2857762
COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION 2983500
COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION -
COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 2122443 2122443

Referencias bibliográficas

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The effect of nicotine on the production of soluble fms-like tyrosine kinase-1 and soluble endoglin in human umbilical vein endothelial cells and trophoblasts.
Ja-Young Kwon,Sang-Wook Bai,Young-Guen Kwon,Se-Hoon Kim,Chul Hoon Kim,Moung Hwa Kang,John A Linton,Yong-Won Park
Acta obstetricia et gynecologica Scandinavica  89  2009

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Casein kinase 1 delta regulates the pace of the mammalian circadian clock.
Jean-Pierre Etchegaray, Kazuhiko K Machida, Elizabeth Noton, Cara M Constance, Robert Dallmann, Marianne N Di Napoli, Jason P DeBruyne, Christopher M Lambert, Elizabeth A Yu, Steven M Reppert, David R Weaver
Molecular and cellular biology  29  3853-66  2009

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The mammalian molecular clockwork controls rhythmic expression of its own input pathway components.
Martina Pfeffer,Christian M Müller,Jérôme Mordel,Hilmar Meissl,Nariman Ansari,Thomas Deller,Horst-Werner Korf,Charlotte von Gall
The Journal of neuroscience : the official journal of the Society for Neuroscience  29  2009

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19439589 19439589
Genotypic variability and persistence of Legionella pneumophila PFGE patterns in 34 cooling towers from two different areas.
Inma Sanchez,Marian Garcia-Nuñez,Sonia Ragull,Nieves Sopena,Maria Luisa Pedro-Botet,Maria Estere,Celestino Rey-Joly,Miquel Sabria,Maria Esteve
Environmental microbiology  10  2008

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18199124 18199124
Prevention of renal cell carcinoma by active vitamin D3.
Fujioka, T, et al.
World journal of surgery, 24: 1205-10 (2000)  1999

Mostrar resumen
11071463 11071463
Rapid colorimetric assay for cell viability: application to the quantitation of cytotoxic and growth inhibitory lymphokines.
Green, L M, et al.
J. Immunol. Methods, 70: 257-68 (1984)  1983

Mostrar resumen
6609997 6609997