Recombinant Botulinum Neurotoxin Type B Heavy Chain was expressed in E. coli.
Clostridium botulinum neurotoxins (BoNT) are among the most toxic substances known to man. Seven immunologically distinct serotypes of neurotoxin, designated types A through G (BoNT/A, BoNT/B, BoNT/C, BoNT/D, BoNT/E, BoNT/F, BoNT/G), have been identified. These botulinum neurotoxins are naturally produced complexed with one or more nontoxic neurotoxin-associated proteins (NAPs). NAPs are important to toxicity by ingestion in that they protect the neurotoxin from proteases, acidity, heat and they may also play a role in translocation across the mucosal layer.
Each vial of recombinant Botulinum Neurotoxin Type B Heavy Chain Binding Domain (HccB) contains 50.0 ug of lyophilized protein. When reconstituted with 500 ul of sterile distilled water, each vial contains 50.0 ug of HccB in 20 mM HEPES, pH 7.4, 1.25% lactose. The protein was recombinantly expressed in E. coli and purified using affinity and anion exchange chromatography. The affinity tag has subsequently been cleaved off of the protein prior to quantitation and packaging.
HccB is highly reactive to Anti-Botulinum Type B antibodies in a Western Blot. Botulinum Neurotoxin Type B Heavy Chain Binding Domain was tested in an ELISA assay using GT1b and the receptor domain of Synaptotagmin II. The midpoint of the binding curve of HccB to Synaptotagmin II was 5.7 ng. The midpoint of the binding curve of HccB to GT1b was 11.4 ng.
HccB is 447 amino acids in length. This product contains amino acids 858-1291 of the Botulinum Neurotoxin Type B Okra Strain. There are also 13 residual amino acids on the N-terminus from the affinity tag. The molecular weight of the protein is approximately 53 kDa.
The protein is greater than 95% pure based on densitometry.
Toxicity: HccB is only a fragment of the Botulinum toxin and is thus non-toxic.
This product is supplied as a lyophilized powder which has been stoppered under vacuum. Reconstitution of the powder should be done with syringe through the rubber stopper to avoid any loss of material. Store lyophilized vials at 4 centigrade. Once dissolved, aliquot and store at -20 centigrade. Refrain from multiple freeze/thaw cycles.
Protein concentration was determined by a modification of the Bradford method using bovine serum albumin as a standard.
Good laboratory technique should be employed in the safe handling of this product. This requlres observing the following practices:
1. Wear appropriate laboratory attire including a lab coat, gloves and safety glasses.
2. Do not mouth pipette, inhale, ingest or allow to come into contact with open wounds. Wash thoroughly any area of the body which comes into contact with the product.
3. Avoid accidental autoinoculation by exercising extreme care when handling in conjunction with any injection device.
4. This product is intended for research purposes by qualified personnel only. It is not intended for use in humans or as a diagnostic agent. Our company is not liable for any damages resulting from the misuse or handling of this product.