Botulinum Neurotoxin Type E was purified from Clostridium botulinum, nicked.
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 10 ug vial of nicked botulinum neurotoxin type E, when reconstituted with 110 ul of water, contains 10 ug of protein in 20 mM sodium citrate, pH 6.0 with 1.25% lactose. This product has been activated by nicking. To ensure full recovery of toxin from the vial, include 1 mg/ml of BSA or 0.2% TWEEN 20 in the reconstitution buffer. Handle the product gently, mix by inversion, do not vortex. READ ALL HANDLING INFORMATION PRIOR TO RECONSTITUTION.
This nicked C. botulinum neurotoxin type E has been tested for activity in an endopeptidase assay using SNAP-25 as the substrate. The neurotoxin, at 20 nM, cleaves >80% of the 5 uM substrate in 15 minutes at 3rC, with complete cleavage in 60 minutes.
When examined on 4-12% SDS-polyacrylamide gels, this product migrates as a single major band with an apparent molecular weight of approximately 150 kDa. In the presence of a reducing agent, the preparation migrates as two major bands corresponding to the heavy chain (100 kDa) and the light chain (50 kDa), respectively. The band corresponding to the intact toxin at 150 kDa is approximately 8% of the total toxin indicating 92% nicked toxin.
The endotoxin content, determined using a kinetic chromagenic LAL assay, is 65.3 EU/mg.
Toxicity: This product has been activated by nicking. Toxicity testing has not been done on this lot. Botulinum toxin is the most deadly bacterial toxin known to man. The minimum lethal dose (L0100) of Clostridium botulinum type E neurotoxin in mice is estimated at 1.1 ng/kg, i.p. Humans are said to be at least as sensitive as mice. 1 Consult the MSOS for further information.
This product is supplied as a lyophilized powder which has been stoppered under vacuum. Store lyophilized vials at 2-8 centigrade.
Good laboratory technique should be employed in the safe handling of this product. This requlres observing the following practices:
1. Persons handling this product and contaminated glassware should consult the current version of the Biosafety in Microbiological and Biomedical Laboratories.
2. This product is to be used by skilled personnel under the direction of a principal investigator in a laboratory setting only.
3. Wear appropriate laboratory attire including a lab coat, gloves and safety glasses.
4. Never remove the stopper prior to reconstitution and never work with the product in the powdered form. Always reconstitute it first.
5. 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.
6. Avoid accidental autoinoculation by exercising extreme care when handling in conjunction with any injection device.
7. 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.