CT02 | MTT Cell Growth Assay Kit

CT02
1000 assays  
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      Overview

      Key Spec Table

      Key Applications
      ACT
      Description
      Catalogue Number CT02
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      Description MTT Cell Growth Assay Kit
      Overview MTT 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.
      References
      Product Information
      Components Solution B: PBS pH 7.4, 15 mL Reagent A: MTT, (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide), 50 mg/vial.
      Applications
      Application MTT Cell Growth Assay is a colorimetric assay that can be used for either proliferation or complement-mediated cytotoxicity assays.
      Key Applications
      • Activity Assay
      Application Notes Procedure:
      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 Conditions Maintain 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 Size 1000 assays
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      MSDS

      Languages
      English

      Certificates of Analysis

      TitleLot Number
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION -
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 2122443 2122443
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 2140033 2140033
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 2329797 2329797
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 2390500 2390500
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 2453245 2453245
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 2465226 2465226
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 1962558 1962558
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 2006356 2006356
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 2030015 2030015
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 2037179 2037179
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 2045849 2045849
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 2075099 2075099
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 2189708 2189708
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 2227132 2227132
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 2266496 2266496
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 2302196 2302196
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 2500654 2500654
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - NG1913429 NG1913429

      References

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      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 2010

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      20367431
      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

      Show Abstract Full Text Article
      19414593
      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
      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
      Prevention of renal cell carcinoma by active vitamin D3.
      Fujioka, T, et al.
      World journal of surgery, 24: 1205-10 (2000) 2000

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      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) 1984

      Show Abstract
      6609997