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322326 Diphtheria Toxin, Unnicked, Corynebacterium diphtheriae - Calbiochem

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322326-1MG
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      Description
      OverviewCatalyzes ADP-ribosylation of eukaryotic aminoacyltransferase II (EF2) using NAD as substrate, thereby inhibiting protein synthesis. May also induce internucleosomal breakdown. Causes DNA fragmentation and cytolysis in U937 cells. Activation requires nicking with a protease followed by reduction with DTT.
      Catalogue Number322326
      Brand Family Calbiochem®
      References
      ReferencesKochi, S.K., and Collier, R.J. 1993. Exp. Cell Res. 208, 296.
      Chang, M.P., et al. 1989. J. Biol. Chem. 264, 15261.
      Pappenheimer, A.M., Jr. 1977. Annu. Rev. Biochem. 46, 69.
      Ittelson, T.R., and Gill, D.M. 1973. Nature 242, 330.
      Uchida, T., et al. 1973. J. Biol. Chem. 248, 3851.
      Pappenheimer, A.M., et al. 1972. Immunochem. 9, 891.
      Uchida, T., et al. 1972. Science 175, 901.
      Bowman, C.G., and Bonventre, P.F. 1970. J. Exp. Med. 131, 659.
      Baseman, J.B., et al. 1970. J. Exp. Med. 132, 1138.
      Gill, D.M., et al. 1969. J. Exp. Med. 129, 1.
      Gill, D.M., et al. 1969. Cold Spring Harbor Symp. Quant. Biol. 34, 589.
      Honjo, J., et al. 1968. J. Biol. Chem. 243, 3553.
      Product Information
      ATP CompetitiveN
      FormSolid
      FormulationLyophilized from sterile 10 mM Tris, 1 mM EDTA, pH 7.5.
      ReversibleN
      Quality LevelMQ100
      Applications
      Biological Information
      Biological activityNote: Toxicity may vary by lot of toxin. Each laboratory should determine the optimum dosage for each particular application.
      Primary TargetEukaryotic aminoacyltransferase II (EF2)
      PurityMajor band (under reducing conditions) of ~63 kDa
      Physicochemical Information
      Cell permeableN
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      RTECSXW5807200
      Safety Information
      R PhraseR: 20/21/22

      Harmful by inhalation, in contact with skin and if swallowed.
      S PhraseS: 36

      Wear suitable protective clothing.
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Ambient Temperature Only
      Toxicity Harmful
      Storage +2°C to +8°C
      Do not freeze Ok to freeze
      Special InstructionsFollowing reconstitution, aliquot, quickly freeze on dry ice, and freeze (-70°C). Subsequent thawing should be carried out only at room temperature. For assays employing very low concentrations of diphtheria toxin, the use of a carrier protein, such as BSA or HSA, is recommended.
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      322326-1MG 07790788060923

      Documentation

      Diphtheria Toxin, Unnicked, Corynebacterium diphtheriae - Calbiochem SDS

      Title

      Safety Data Sheet (SDS) 

      Diphtheria Toxin, Unnicked, Corynebacterium diphtheriae - Calbiochem Certificates of Analysis

      TitleLot Number
      322326

      References

      Reference overview
      Kochi, S.K., and Collier, R.J. 1993. Exp. Cell Res. 208, 296.
      Chang, M.P., et al. 1989. J. Biol. Chem. 264, 15261.
      Pappenheimer, A.M., Jr. 1977. Annu. Rev. Biochem. 46, 69.
      Ittelson, T.R., and Gill, D.M. 1973. Nature 242, 330.
      Uchida, T., et al. 1973. J. Biol. Chem. 248, 3851.
      Pappenheimer, A.M., et al. 1972. Immunochem. 9, 891.
      Uchida, T., et al. 1972. Science 175, 901.
      Bowman, C.G., and Bonventre, P.F. 1970. J. Exp. Med. 131, 659.
      Baseman, J.B., et al. 1970. J. Exp. Med. 132, 1138.
      Gill, D.M., et al. 1969. J. Exp. Med. 129, 1.
      Gill, D.M., et al. 1969. Cold Spring Harbor Symp. Quant. Biol. 34, 589.
      Honjo, J., et al. 1968. J. Biol. Chem. 243, 3553.
      Data Sheet

      Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

      Revision14-July-2017 JSW
      DescriptionCatalyzes ADP-ribosylation of eukaryotic aminoacyltransferase II (EF2) using NAD as substrate, thereby inhibiting protein synthesis. May also induce internucleosomal breakdown. Causes DNA fragmentation and cytolysis in U937 cells. Activation requires nicking with a protease followed by reduction with DTT. Diphtheria toxin, secreted by certain strains of Corynebacterium diphtheriae, catalyzes the ADP-ribosylation of eukaryotic aminoacyl transferase II (EF2) using NAD as a substrate. The reaction is the basis of its toxicity towards eukaryotic organisms.

      Diphtheria toxin, synthesized and excreted as a proenzyme, is composed of a single polypeptide chain of 63 kDa. For its enzymatic activity to be expressed, the toxin must undergo two covalent alterations in structure. First, a mild proteolysis results in the formation of an enzymatically inactive "nicked toxin," which consists of two major fragments (A and B) linked by a disulfide bond. Reduction of the nicked toxin with thiols (DTT) releases the enzymatically active N-terminal A fragment (24 kDa). The C-terminal B fragment (39 kDa) has no apparent enzymatic activity. The B fragment is required for toxicity and is responsible for recognizing and binding of the toxin to cell surface receptors. Diphtheria toxin is purified from Corynebacterium diphtheriae Park Williams strain 8 by a modified method of Pappenheimer, et al. As assessed by disc electrophoresis run at alkaline pH under non-denaturing conditions, this preparation migrates as a major band at 63 kDa, corresponding to the intact toxin. Two faster, more lightly stained bands (24 and 39 kDa), corresponding to A and B fragments, may be observed. Following trypsin treatment in DTT, diphtheria toxin exhibits high activity when assayed for its ability to ADP-ribosylate EF2. The ED50 for CHO cells is determined to be about 0.4 ng/ml.
      FormSolid
      FormulationLyophilized from sterile 10 mM Tris, 1 mM EDTA, pH 7.5.
      RTECSXW5807200
      PurityMajor band (under reducing conditions) of ~63 kDa
      Biological activityNote: Toxicity may vary by lot of toxin. Each laboratory should determine the optimum dosage for each particular application.
      SolubilityAqueous buffers or H₂O. Please see vial label for lot-specific reconstitution volume.
      Storage +2°C to +8°C
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing reconstitution, aliquot, quickly freeze on dry ice, and freeze (-70°C). Subsequent thawing should be carried out only at room temperature. For assays employing very low concentrations of diphtheria toxin, the use of a carrier protein, such as BSA or HSA, is recommended.
      Toxicity Harmful
      ReferencesKochi, S.K., and Collier, R.J. 1993. Exp. Cell Res. 208, 296.
      Chang, M.P., et al. 1989. J. Biol. Chem. 264, 15261.
      Pappenheimer, A.M., Jr. 1977. Annu. Rev. Biochem. 46, 69.
      Ittelson, T.R., and Gill, D.M. 1973. Nature 242, 330.
      Uchida, T., et al. 1973. J. Biol. Chem. 248, 3851.
      Pappenheimer, A.M., et al. 1972. Immunochem. 9, 891.
      Uchida, T., et al. 1972. Science 175, 901.
      Bowman, C.G., and Bonventre, P.F. 1970. J. Exp. Med. 131, 659.
      Baseman, J.B., et al. 1970. J. Exp. Med. 132, 1138.
      Gill, D.M., et al. 1969. J. Exp. Med. 129, 1.
      Gill, D.M., et al. 1969. Cold Spring Harbor Symp. Quant. Biol. 34, 589.
      Honjo, J., et al. 1968. J. Biol. Chem. 243, 3553.