Recombinant Tetanus Neurotoxin Type Light Chain Protein, CF

R&D Systems | Catalog # 6535-ZN

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Tetanus Neurotoxin Type Light Chain Protein (6535-ZN)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

E. coli

Accession Number

P04958

Applications

Enzyme Activity
View all TeNT Light Chain Proteins and Enzymes »

Product Specifications

Source

E. coli-derived c. tetani TeNT Light Chain protein
Pro2-Gly430, with an N-terminal Met and 6-His tag

Purity

>85%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

50 kDa

SDS-PAGE

43-50 kDa, reducing conditions

Activity

Measured by the cleavage of the substrate GFPuv/SNAP/VAMP in a gel-shift assay.
>50% of 1 μg substrate is cleaved by 40 ng, as measured under the described conditions.

Formulation, Preparation, and Storage

6535-ZN
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Tween® 20.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: TeNT Light Chain

Tetanus toxin (TeNT) is a potent neurotoxin produced by Clostridium tetani. TeNT is similar in structure and function to the botulinum toxins produced by other Clostridium species (1, 2). TeNT is among the most toxic protein toxins known for humans. TeNT is synthesized as an inactive single chain precursor that undergoes proteolytic cleavage to create light and heavy chains that are linked by a disulfide bond. The 50 kDa light chain contains a metalloprotease domain whereas the 100 kDa heavy chain contains a receptor binding domain and a domain required for translocation across the cell membrane (3). Once inside a neuronal cell the zinc metalloprotease domain is able to cleave synaptobrevin to cause motor neuron disinhibition, resulting in the spastic paralysis characteristic of tetanus (4). The E. coli expressed recombinant TeNT light chain is an active protease. In the absence of heavy chains, however, it lacks toxicity because it cannot enter into host cells.

References

  1. Eisel, U. et al. (1986) EMBO J. 5:2495.
  2. Montecucco, C. and S. Giampietro (1993) Trends Biochem. Sci. 18:324.
  3. Schiavo, G. et al. (1992) Nature 359:832.
  4. Schiavo, G. et al. (1992) EMBO J. 11:3577.

Long Name

Tetanus Neurotoxin Type Light Chain

Alternate Names

TeNT-LC, Tentoxylysin, tetX

Entrez Gene IDs

1061100 (C. tetani)

Gene Symbol

TETX

UniProt

P04958 (C. tetani)

Additional TeNT Light Chain Products

Product Documents for Recombinant Tetanus Neurotoxin Type Light Chain Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Tetanus Neurotoxin Type Light Chain Protein, CF

Coomassie is a registered trademark of Imperial Chemical Industries Ltd. Tween is a registered trademark of ICI Americas.

For research use only

Related Research Areas

Metalloproteases and Regulators

Citations for Recombinant Tetanus Neurotoxin Type Light Chain Protein, CF

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Protocols

View specific protocols for Recombinant Tetanus Neurotoxin Type Light Chain Protein, CF (6535-ZN):

Materials
  • Assay Buffer: 50 mM HEPES, 0.05% (v/v) Tween® 20, pH 7.5
  • Recombinant C. tetani TeNT Light Chain (rTeNT-LC) (Catalog # 6535-ZN)
  • Substrate: Recombinant GFP/SNAP25B/VAMP-2 (Catalog # 7375-SV)
  • SDS-PAGE and silver stain reagents or equivalent or Western blot with appropriate antibodies
  1. Dilute Substrate to 100 µg/mL in Assay Buffer.
  2. Dilute rTeNT-LC to 4 µg/mL in Assay Buffer.
  3. Combine equal volumes of diluted Substrate with diluted rTeNT-LC. Prepare two controls by combining equal volumes of diluted Substrate with Assay Buffer.
  4. Incubate reaction vials at 37 °C for 1 hour.  Incubate one control at 37 °C and the other at -20 °C for 1 hour.
  5. After incubation, combine reaction mixtures and controls with reducing SDS-PAGE sample buffer at a 1:1 (reaction mixture:sample buffer) ratio (v/v) to stop reactions.
  6. Analyze the cleavage products by SDS-PAGE (Load 40 µL of the mixture from step 5 per lane, 1 µg Substrate per lane) followed by silver staining and/or Western blot.
  • rTeNT-LC:  40 ng
  • Substrate:  1 µg

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