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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).

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
567333
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Overview

Replacement Information

Products

Catalogue NumberPackaging Qty/Pack
567333-200NMOL Plastic ampoule 200 nmol
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.