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CBA055 K-LISA™ mTOR Activity Kit

K-LISA™ mTOR Activity Kit MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
CBA055
  
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      Overview

      Key Spec Table

      Detection Methods
      Colorimetric
      Description
      Overview

      This product has been discontinued.



      An ELISA-based activity assay for measuring the kinase activity of purified or mTOR immunoprecipitated from cell lysates. Useful for in vitro mTOR inhibitor screening and for assessing the regulation of mTOR cell signaling. This assay utilizes a p70S6K GST fusion protein as a specific mTOR substrate.
      Catalogue NumberCBA055
      Brand Family Calbiochem®
      Materials Required but Not Delivered• Tris-buffered Saline (TBS)
      • Wash bottle or multichannel dispenser for washing
      • Protein assay (for cell lysates; e.g. BCA Protein Assay Kit, Cat. No. 71285 or Non-Interfering Protein Assay™ Kit, Cat. No. 488250)
      • Anti-mTOR/FRAP (Ab-2) Mouse mAb (22C2) (Cat. No. OP97) (optional; for immunoprecipitation of mTOR from crude lysates)
      • Protease inhibitor cocktail (e.g. Protease Inhibitor Cocktail Set III, Cat. No. 539134)
      • Protein A or Protein G agarose for immunoprecipitation of mTOR from cell lysates (e.g. Protein A Agarose Suspension, Cat. No. IP02; Protein G-PLUS Agarose Suspension, Cat. No. IP04; or Protein G-PLUS/ Protein A Agarose Suspension, Cat. No. IP05)
      • Spectrophotometer capable of measuring absorbance in 96-well plates at 450 nm, preferably in the dual wavelength mode at 450 nm against a reference wavelength of 540 or 595 nm.
      References
      ReferencesKong, D. and Yamori, T. 2007. Cancer Sci., 98, 1638.
      Mavarakis, M., et al. 2006. Human Mol Genetics 15, 2962.
      Chiang G.G. and Abraham R.T. 2005. J. Biol. Chem. 280, 25485.
      Guertin, D.A., et al. 2005. Trends Mol. Med. 11, 353.
      Sarbassov, D.D., et al. 2005. Science 307, 1098.
      Chiang, G.G. and Abraham, R.T. 2004. Methods Mol. Biol. 281, 125.
      Sarbassov, D.D., et al. 2004. Curr. Biol. 14, 1296.
      Kimura, N., et al. 2003. Genes Cells 8, 65.
      Kim, D.H., et al. 2002. Cell 110, 163.
      Sekulic, A. et al. 2000. Cancer Res. 60, 3504.
      Brunn, G.J., et al. 1997. Science 277, 99.
      Alarcon, C.M., et al. 1996. Genes Dev. 10, 279.
      Brunn, G.J., et al. 1996. EMBO J. 15, 5256.
      Freeman, K. and Livi, G.P. 1996. Gene 172, 143.
      Zheng, X.F., et al. 1995. Cell 82, 121.
      Brown, E.J., et al. 1994. Nature 369, 756.
      Sabatini, D.M., et al. 1994. Cell 78, 35.
      Stan, R., et al. 1994. J. Biol. Chem. 269, 32027.
      Heitman, J., et al. 1991. Science 253, 905.
      Product Information
      Detection methodColorimetric
      DeclarationSold under license of U.S. Patent 7,135,298.
      Form96 Tests
      Format96-well plate
      Kit containsGlutathione-Coated 96-Well Plate, mTOR Substrate, Anti-p70S6K-T389, mTOR Standard, 2X Kinase Assay Buffer, ATP Solution, 100 mM DTT, Wortmannin 50X, Kinase Stop Solution, 10X Plate Wash Concentrate, Antibody Diluent, HRP-Antibody Conjugate, TMB Substrate, ELISA Stop Solution, Plate Sealers and a user protocol.
      Quality LevelMQ100
      Biological Information
      Sample TypePurified or partially purified enzyme
      Product Usage Statements
      Intended useThe K-LISA™ mTOR Activity Kit is suitable for measuring the kinase activity of purified or partially purified preparations of mTOR or mTOR that has been immunoprecipitated from crude cell lysates. The assay is also useful for in vitro mTOR inhibitor screening and for assessing the regulation of mTOR in cell signaling.
      Storage and Shipping Information
      Ship Code Dry Ice Only
      Toxicity Multiple Toxicity Values, refer to MSDS
      Storage ≤ -70°C
      Storage ConditionsUpon arrival store the Glutathione-Coated 96-Well Plate, the Kinase Stop Solution, and the Anti-p70S6K-T389 at 4°C; store the mTOR Standard, mTOR Substrate, 100 mM DTT, and 20 mM ATP Solution at -70°C; store the remaining components of the kit at -20°C.
      Note: THE KIT IS SHIPPED ON DRY ICE, BUT SOME COMPONENTS ARE STORED AT 4°C FOR LONG-TERM STORAGE. DO NOT FREEZE/THAW THE ANTI-p70S6K-T389 ANTIBODY
      Avoid freeze/thaw Avoid freeze/thaw
      Do not freeze Ok to freeze
      Supplemental Information
      Kit containsGlutathione-Coated 96-Well Plate, mTOR Substrate, Anti-p70S6K-T389, mTOR Standard, 2X Kinase Assay Buffer, ATP Solution, 100 mM DTT, Wortmannin 50X, Kinase Stop Solution, 10X Plate Wash Concentrate, Antibody Diluent, HRP-Antibody Conjugate, TMB Substrate, ELISA Stop Solution, Plate Sealers and a user protocol.
      Global Trade Item Number
      Catalogue Number GTIN
      CBA055 0