567333 SNAPtide® Botulinum Toxin A Substrate, Fluorogenic - Calbiochem

567333
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      Overview

      Replacement Information

      Pricing & Availability

      Catalog Number AvailabilityPackaging Qty/Pack Price Quantity
      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
          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

          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.