Botulinum Neurotoxin Type A, Light Chain (BoNT/A LC)

Size

£843.00

SKU:: 215914
Purity:: Purified
Storage Conditions:: -20°C
Supplier:: United States Biological
Host:: Mouse
Immunogen:: Botulinum Neurotoxin Type A, Light Chain.
Clone:: F1-40
Format:: Supplied as a liquid in PBS, pH 7.4, 0.05% sodium azide
Extra Details:: The potent toxicity of both the botulinum neurotoxins and anthrax lethal toxin is due to a zinc- dependent proteolytic activity associated with the toxins. Measurement of this enzymatic activity provides for both a potentially sensitive and direct means for detection of the toxin, and a method for identifying potential toxin inhibitors using high throughput screening. A highly efficient approach for monitoring enzymatic activity is based on the use of fluorescence resonance energy transfer (FRET) substrates. These fluorogenic peptides contain a donor fluorescent group at one end and a suitable chromogenic acceptor group at the other. The fluorescence is quenched initially by intramolecular energy transfer between the donor/acceptor pair. Cleavage of the FRET substrate by the appropriate enzyme releases the fluorophore and full fluorescence is restored. The increase in fluorescence intensity is directly proportional to the amount of enzyme present. Enzymatic activity can be monitored continuously by recording the increase in fluorescence intensity with time. The change in the relative fluorescence units (RFU) as cleavage occurs can be converted to nmoles of cleaved substrate from a standard curve generated using a Calibration Peptide which is the cleaved substrate containing only the N-terminally attached fluorophore. For Botulinum neurotoxin type A, a Control FRET peptide substrate that is not cleaved by the neurotoxin but contains all remaining non-specific sites in the sequence can be used to screen background cleavage of the substrate that can occur in complex matrices. There are seven immunologically distinct botulinum neurotoxins, designated types A-G. Each neurotoxin toxin cleaves one of two target proteins critical for synaptic vesicle fusion to the plasma membrane and neurotransmitter release, either SNAP-25 or Synaptobrevin. The latter is also called VAMP-2. Types A, C and E specifically bind and selectively cleave SNAP-25 while types B, D, F and G cleave Synaptobrevin. Type C also cleaves Syntaxin. The enzymatic activity of these toxins can also be measured using the appropriate native substrate and monitoring the generation of the cleaved product on SDS-PAGE gel electrophoresis. The synaptic vesicle glycoprotein 2c (SV2c) has been shown to be the protein receptor for Botulinum neurotoxin, Type A (BoNT/A) and the luminal domain loop is the specific location of BoNT/A binding. Applications: Suitable for use in Sandwich ELISA and Dot Blot. Other applications have not been tested. Recommended Dilutions: Dot Blot: 1.25ug/ml solution of the antibody was an effective probe for 0.05ug of BoNT/A. ELISA (Sandwich): Can be used as a capture antibody and paired with 215913 to detect BoNT/A Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Shipping Conditions:: Blue Ice