Home / BoNT-E Heavy Chain / C. botulinum BoNT-E Heavy Chain Antibody

C. botulinum BoNT-E Heavy Chain Antibody

Catalog #: MAB7135 Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
MAB7135
MAB7135-SP
Detection ofC. botulinumBoNT‑E Heavy Chain by Western Blot.
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C. botulinum BoNT-E Heavy Chain Antibody Summary

Species Reactivity
C. botulinum
Specificity
Detects C. botulinum BoNT-E Heavy Chain in direct ELISAs and Western blots. In direct ELISAs, no cross-reactivity with recombinant Heavy Chains from BoNT-A or BoNT-G is observed.
Source
Monoclonal Mouse IgG2B Clone # 785208
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
E. coli-derived recombinant C. botulinum BoNT-E Heavy Chain
Lys845-Lys1251
Accession # P30995
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of<EM>C. botulinum</EM>BoNT-E Heavy Chain antibody by Western Blot. View Larger

Detection ofC. botulinumBoNT‑E Heavy Chain by Western Blot. Western blot shows lysates of recombinantC. botulinumBoNT-E Heavy Chain Receptor Binding Domain (25 ng/lane). PVDF membrane was probed with 1 µg/mL of Mouse Anti-C. botulinumBoNT-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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.5 mg/mL.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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 Clostridium botulinum (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.

References
  1. Campbell K.D. et al. (1993) J. Clin. Microbiol. 31:2255.
  2. Montecucco, C. and Giampietro, S. (1993) Trends. Biochem. Sci. 18:324.
  3. Turton, K. et al. (2002) Trends. Biochem. Sci. 27:552.
  4. Schiavo, G. et al. (2000) Physiol. Rev. 80:717.
Long Name
Botulinum Neurotoxin Type E Heavy Chain
Alternate Names
BoNTE Heavy Chain; BoNT-E Heavy Chain; Bontoxilysin-E

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