CT01 MTT Cell Growth Assay Kit

CT01
5000 assays  
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      Overview

      Replacement Information

      Key Specifications Table

      Key Applications
      ACT
      Description
      Catalogue NumberCT01
      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, 60 mL
      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:
      For every 1,000 assays to be performed, 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 Size5000 assays
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number
      COLORIMETRIC (MTT) KIT2470840
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION2476974
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION2970688
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION2950708
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION2992124
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION2908126
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION3082338
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION2868012
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION2928813
      COLORIMETRIC (MTT) KIT FOR CELL SURVIVAL AND PROLIFERATION - 24363942436394

      References

      Reference overviewPub Med ID
      Ovalicin ameliorates compound 48/80-induced atopic dermatitis-related symptoms.
      Cheol-Sik Yoon,Sung-Hee Nam,Ji-Young Jeon,Hei-Sam Lee,Myeong-Lyeol Lee,Hyeong-U Son,Sang-Han Lee
      Biological & pharmaceutical bulletin  34  2011

      Show Abstract
      22130246 22130246
      Discovery of a novel class of covalent inhibitor for aldehyde dehydrogenases.
      May Khanna,Che-Hong Chen,Ann Kimble-Hill,Bibek Parajuli,Samantha Perez-Miller,Sulochanadevi Baskaran,Jeewon Kim,Karl Dria,Vasilis Vasiliou,Daria Mochly-Rosen,Thomas D Hurley
      The Journal of biological chemistry  286  2011

      Show Abstract
      22021038 22021038
      Characterization of a novel rat cholangiocarcinoma cell culture Model-CGCCA.
      Yeh CN, Lin KJ, Chen TW, Wu RC, Tsao LC, Chen YT, Weng WH, Chen MF
      World journal of gastroenterology : WJG  17  2924-32.  2011

      Full Text Article
      21734803 21734803
      Cobalt chloride decreases EC-SOD expression through intracellular ROS generation and p38-MAPK pathways in COS7 cells.
      Tetsuro Kamiya,Hirokazu Hara,Harutaka Yamada,Hirokazu Imai,Naoki Inagaki,Tetsuo Adachi
      Free radical research  42  2008

      Show Abstract
      19031313 19031313
      Identification of genes required for recycling reducing power during photosynthetic growth.
      Christine L Tavano,Angela M Podevels,Timothy J Donohue
      Journal of bacteriology  187  2005

      Show Abstract Full Text Article
      16030219 16030219
      I. Poloxamer-formulated plasmid DNA-based human cytomegalovirus vaccine: evaluation of plasmid DNA biodistribution/persistence and integration.
      Adrián Vilalta,Rohit K Mahajan,Jukka Hartikka,Denis Rusalov,Terrie Martin,Vesselina Bozoukova,Vicky Leamy,Keith Hall,Peggy Lalor,Alain Rolland,David C Kaslow
      Human gene therapy  16  2005

      Show Abstract
      16218775 16218775
      Prevention of renal cell carcinoma by active vitamin D3.
      Fujioka, T, et al.
      World journal of surgery, 24: 1205-10 (2000)  2000

      Show Abstract
      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)  1984

      Show Abstract
      6609997 6609997

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