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567333 SNAPtide® Botulinum Toxin A Substrate, Fluorogenic - Calbiochem

SNAPtide® Botulinum Toxin A Substrate, Fluorogenic, is a synthetic peptide that acts as a FRET substrate with the fluorophore o-aminobenzoic acid and the acceptor chromophore 2,4-dinitrophenol (DNP).

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567333
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Overview

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567333-200NMOL
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      		 Plastic ampoule 200 nmol
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      Description
      OverviewA synthetic peptide, fluorescence resonance energy transfer (FRET) substrate containing the N-terminally-linked fluorophore o-aminobenzoic acid (Abz) and the acceptor chromophore 2,4-dinitrophenol (DNP) linked to the C-terminus. The substrate is based on the native botulinum toxin type A cleavage site found in the synaptosome-associated protein SNAP-25. Cleavage of the substrate by botulinum toxin releases the fluorophore and full fluorescence is restored.
      Catalogue Number567333
      Brand Family Calbiochem®
      References
      ReferencesBigalke, H. and Shoer, L.F. 2000. In: Handbook of Experimental Pharmacology, 145, Bacterial Protein Toxins (K. Aktories and I. Just, Eds.) pp. 407-443, Springer-Verlag, Berlin.
      Schiavo, G., et al. 1993. J. Biol. Chem. 268, 23784.
      Product Information
      ATP CompetitiveN
      FormLyophilized
      FormulationLyophilized from water.
      ReversibleN
      Quality LevelMQ100
      Applications
      Biological Information
      Primary TargetSubstrate for the native botulinum toxin
      Purity≥90% by HPLC
      Physicochemical Information
      Cell permeableN
      Emission max.
      Excitation max.
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Ambient Temperature Only
      Toxicity Standard Handling
      Storage +2°C to +8°C
      Do not freeze Ok to freeze
      Special InstructionsFollowing reconstitution, aliquot, cover with foil to protect from light, and freeze (-20°C). Stock solutions are stable for up to 3 months at -20°C.
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      567333-200NMOL 04055977267211

      Documentation

      SNAPtide® Botulinum Toxin A Substrate, Fluorogenic - Calbiochem SDS

      Title

      Safety Data Sheet (SDS) 

      SNAPtide® Botulinum Toxin A Substrate, Fluorogenic - Calbiochem Certificates of Analysis

      TitleLot Number
      567333

      References

      Reference overview
      Bigalke, H. and Shoer, L.F. 2000. In: Handbook of Experimental Pharmacology, 145, Bacterial Protein Toxins (K. Aktories and I. Just, Eds.) pp. 407-443, Springer-Verlag, Berlin.
      Schiavo, G., et al. 1993. J. Biol. Chem. 268, 23784.
      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.

      Revision28-February-2012 JSW
      DescriptionA synthetic peptide, fluorescence resonance energy transfer (FRET) substrate containing the N-terminally-linked fluorophore o-aminobenzoic acid (Abz) and the acceptor chromophore 2,4-dinitrophenol (DNP) linked to the C-terminus. The substrate is based on the native botulinum toxin type A cleavage site found in the synaptosome-associated protein SNAP-25. Cleavage of the substrate by botulinum toxin releases the fluorophore and full fluorescence is restored.
      FormLyophilized
      FormulationLyophilized from water.
      Recommended reaction conditionsThe FRET assays are performed using HEPES buffers prepared by titrating the free acid form of HEPES with the potassium salt form of HEPES. For assays with BoNT/A, the SNAPtide® stock solution is diluted using 20 mM HEPES, pH 8.0, prior to use. For assays with BoNT/A Light Chain, the stock solution should be diluted in the hydrolysis buffer, described in the section below. When using a 96-well plate and a final volume of 250 µl/well, a 250 µM stock solution is convenient to use. The final concentration of SNAPtide® to be used is typically between 5 µM and 10 µM/well, depending on the instrumentation and experiment. Since DMSO inhibits cleavage, final concentrations must be less than 2% of the total volume. For SNAPtide® (o-Abz/Dnp any concentration of ZnCl2 in the BoNT/A Light Chain hydrolysis buffer inhibits cleavage.

      These FRET assays are run at 37°C. Excitation wavelength is 320 nm and emission is 420 nm. There is a linear dependence of fluorescence intensity on concentration of totally cleaved substrate up to 30 ~M SNAPtide®(o-Abz/Dnp). When measuring kinetic parameters such as the Km and Vmax for this FRET substrate, the data must be corrected for a phenomenon known as the "inner filter effect". This effect, as well as a method to determine an appropriate correction factor, is explained in the paper by Liu, et al. 1999. Analytical Biochemistry 267, 331. The correction method uses an unquenched fluorophore for comparison.
      Purity≥90% by HPLC
      SolubilityDMSO (5 mM). Prepare a 5 mM stock solution of this peptide in DMSO as follows: Add 40 µl of DMSO to a vial containing 200 nmol of peptide.
      Storage +2°C to +8°C
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing reconstitution, aliquot, cover with foil to protect from light, and freeze (-20°C). Stock solutions are stable for up to 3 months at -20°C.
      Toxicity Standard Handling
      ReferencesBigalke, H. and Shoer, L.F. 2000. In: Handbook of Experimental Pharmacology, 145, Bacterial Protein Toxins (K. Aktories and I. Just, Eds.) pp. 407-443, Springer-Verlag, Berlin.
      Schiavo, G., et al. 1993. J. Biol. Chem. 268, 23784.