Detection of C. botulinum BoNT‑E Heavy Chain by Western Blot.
Western blot shows lysates of recombinant C. botulinum BoNT-E Heavy Chain (25 ng/lane). PVDF membrane was probed with 1 µg/mL of Mouse Anti-C. botulinum BoNT‑E Heavy Chain Monoclonal Antibody (Catalog # MAB7135) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for BoNT‑E Heavy Chain at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: BoNT-E Heavy Chain
Botulinum Neurotoxin Type E is one of the seven serotypes of Botulinum Neurotoxins (BoNTs) produced by various strains of Clostridiumbotulinum (1, 2). BoNTs are synthesized as inactive single chain protein precursors and activated by proteolytic cleavage to generate disulfide-linked two-chain proteins. The 50 kDa light chain contains the catalytic domain, whereas the 100 kDa heavy chain contains an internal translocation domain and a receptor binding domain (3). BoNTs are the most potent protein toxins for humans. As zinc proteases, they cleave SNARE proteins to elicit flaccid paralysis in botulism by blocking acetylcholine release at the neuromuscular junction (2‑4). E. coli-expressed recombinant light chains are active proteases. In the absence of the heavy chains, however, they lack toxicity because they cannot enter into host cells.
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Montecucco, C. and Giampietro, S. (1993) Trends. Biochem. Sci. 18:324.
Turton, K. et al. (2002) Trends. Biochem. Sci. 27:552.
Schiavo, G. et al. (2000) Physiol. Rev. 80:717.
Botulinum Neurotoxin Type E Heavy Chain
BoNTE Heavy Chain; BoNT-E Heavy Chain; Bontoxilysin-E
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