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CT01-5 MTT Reagent A

Useful in the study of cell & mitochondrial health, MTT is a pale yellow substrate that is cleaved by living cells to yield a dark blue formazan product.

MTT Reagent A MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Key Applications: Activity Assay
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CT01-5
5 vials  
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      Overview

      Replacement Information

      Key Spec Table

      Key Applications
      ACT
      Description
      Catalogue NumberCT01-5
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      DescriptionMTT Reagent A
      OverviewReagent A: MTT, (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide), 50mg/vial.

      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. Reagent A is a component of Chemicon's MTT assay kit, Catalog Number CT01/CT02.
      Materials Required but Not Delivered• Solution B (not provided): PBS pH 7.4, 60 mL
      • Solution C (not provided): Color development solution (isopropanol with 0.04 N HCl), 500 mL
      References
      Product Information
      Quality LevelMQ100
      Applications
      ApplicationUseful in the study of cell & mitochondrial health, MTT is a pale yellow substrate that is cleaved by living cells to yield a dark blue formazan product.
      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 Solution C 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 l x 106 cells/mL.
      Biological Information
      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 Size5 vials
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      CT01-5 04053252274145

      Documentation

      MTT Reagent A SDS

      Title

      Safety Data Sheet (SDS) 

      MTT Reagent A Certificates of Analysis

      TitleLot Number
      REAGENT A FROM COLORIMETRIC (MTT) ASSAY FOR CELL SURVIVAL AND PROLIFERATION - 2120953 2120953
      REAGENT A FROM COLORIMETRIC (MTT) ASSAY FOR CELL SURVIVAL AND PROLIFERATION - 2139986 2139986
      REAGENT A FROM COLORIMETRIC (MTT) ASSAY FOR CELL SURVIVAL AND PROLIFERATION - 2145123 2145123
      REAGENT A FROM COLORIMETRIC (MTT) ASSAY FOR CELL SURVIVAL AND PROLIFERATION - 2445844 2445844
      REAGENT A FROM COLORIMETRIC (MTT) ASSAY FOR CELL SURVIVAL AND PROLIFERATION - 2453140 2453140
      REAGENT A FROM -2519298 2519298
      REAGENT A FROM -2578092 2578092
      REAGENT A FROM -2689131 2689131
      REAGENT A FROM -2707828 2707828
      REAGENT A FROM -2736659 2736659
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      References

      Reference overviewPub Med ID
      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

      Data Sheet

      Title
      REAGENT A FROM COLORIMETRIC (MTT) ASSAY FOR CELL SURVIVAL AND PROLIFERATION