TEV Protease-Labelled Magnetic Nanoparticles

Tobacco Etch Virus protease (TEVp)-Streptavidin fusion protein immobilized on biotin-coated superparamagnetic nanoparticles for optimal recovery and reuse in protein preparation applications.

Highlights:

  • Significantly reduces downstream protease contamination
  • Simplifies protein preparations by abolishing the need to remove TEVp via column chromatography
  • Active in multiple buffer systems (e.g., PBS; TBS; Imidazole)
  • Suitable for cleavage of affinity tags, identification of TEV sites in proteins, and activation/inactivation of engineered proteins
  • Beads can be reused for multiple protease reactions

Proteases with highly specific activities have applications in the purification and downstream processing of overexpressed proteins, including the cleavage of affinity tags and solubility promoting partners such as GST and MBP. However, proteases can be challenging to express and purify, and commercially sourced proteases such as TEV and 3C can be prohibitively costly.

From the laboratory of Robert M. Hughes, PhD, East Carolina University.

Catalog Number Product Size AVAILABILITY Price Qty
ECU016
TEV Protease-Labelled Magnetic Nanoparticles, 2.5mg
2.5mg 2 weeks
Price: $225.00
Specifications
Product Type: Small Molecule
Name: TEV Protease-Labelled Magnetic Nanoparticles
Core: Magnetic bead
Functionalization: Biotin coated NPs are loaded with TEVp-Streptavidin fusion protein
Format: Liquid
Stability: TEV beads gradually lose activity in cold storage. Activity can be readily restored with the addition of reductant to the proteolysis reaction activity upon reactivation with reductant after storage for two weeks at +4C
Concentration: 5 mg/mL
Mean Diameter: 0.5 microns
Comments: End-users who obtain this product will need a magnet for bead separation
Storage: +4C (do not freeze)
Shipped: Cold packs
Research
Data

(Left) SDSPAGE Analysis of TEV Bead Cleavage of a MBP-ENLYFQS-GFP Substrate Protein at 4 °C and 30 °C. 200 μL of a 74.6 kD substrate protein (0.1 mg/mL) is efficiently cleaved (90%) after overnight incubation with 50 μL of TEV beads at 30 °C. By contrast, overnight incubation at 4 °C results in significantly less efficient cleavage (21%). (Right) TEV beads remain active for at least one month in cold storage (4 °C ). Gradient SDS-PAGE gel shows test sample cleavage after overnight incubation. 200 uL of a 74.6 kD substrate protein (0.1 mg/mL) is efficiently cleaved (90%) after overnight incubation with 50 uL of TEV beads at 30 °C.
Provider
From the laboratory of Robert M. Hughes, PhD, East Carolina University.
Comments

Suggested reaction conditions: Utilize 50-100 μL bead suspension per 250 μL-1 mL reaction; conduct reactions at 30-34C for 6 h to overnight on rotator. Collect beads on magnet for 5 min prior to recovery of supernatant. Beads are active for > 1 month if stored at 4C. Beads can be reused for multiple protease reactions.

Tip: Add DTT or β-ME to a final concentration of 10 mM for optimal substrate proteolysis.

References

If you publish research with this product, please let us know so we can cite your paper.

 
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