Recombinant Mouse Furin Protein, CF

R&D Systems | Catalog # 6450-SE

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Key Product Details

  • R&D Systems CHO-derived Recombinant Mouse Furin Protein (6450-SE)
  • 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

CHO

Accession Number

P23188

Applications

Enzyme Activity
View all Furin Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived mouse Furin protein
Gln25-Glu714, with a C-terminal 10‑His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

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

N-terminal Sequence Analysis

Asp108, Cys198, Ser239, Glu684

Predicted Molecular Mass

77 kDa

SDS-PAGE

60-80 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate pERTKR-AMC (Catalog # ES013).
The specific activity is >150 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

6450-SE
Formulation Supplied as a 0.2 μm filtered solution in Tris, CaCl2, NaCl, Brij-35 and Glycerol.
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: Furin

Furin is a member of the proprotein convertase (PC) family, which belongs to the subtilisin superfamily of serine proteases. As a cellular protease, Furin processes a variety of proproteins in secretory pathway compartments by cleaving after Arg‑Xaa‑Lys/Arg‑Arg‑like motifs, which usually reside at the end of the pro regions of these proproteins. Examples of the proprotein substrates are growth factors and receptors, extracellular matrix proteins, and other proteases. Furin has an essential role in embryogenesis and homeostasis and is implicated in various pathologies such as cancer, neurodegenerative diseases and anthrax (1, 2). Mouse Furin is a 793 amino acid type I transmembrane protein precursor with a signal peptide (residues 1‑24), a pro region (residues 25‑107), which play a crucial role in the folding, activation and transport of Furin, and a mature chain (residues 108‑793) (2, 3). Mouse Furin displays ~94% homology to the human Furin sequence and >99% homology to the subtilisin‑like catalytic domain (3). The mature chain consists of the subtilisin‑like catalytic domain, a P domain, which is essential for enzyme activity and the modulation of pH and calcium requirements, and a cytoplasmic domain, which controls the localization and sorting of Furin in the trans‑Golgi network/endosomal system (1). The purified recombinant mouse Furin (residues 108‑714) corresponds to the mature enzyme terminated before the transmembrane domain.

References

  1. Thomas, G. (2002) Nature Rev. Mol. Cell Biol. 3:753.
  2. Creemers, J.W. and W.J. Van de Ven. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.), pp. 1531, Academic Press, San Diego.
  3. Hatsuzawa, K. et al. (1990) J. Biol. Chem. 265:22075.

Alternate Names

FUR, PACE, PCSK3, SPC1

Entrez Gene IDs

5045 (Human); 18550 (Mouse)

Gene Symbol

FURIN

UniProt

P23188

Additional Furin Products

Product Documents for Recombinant Mouse Furin Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Mouse Furin Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Mouse Furin Protein, CF (6450-SE):

Materials
  • Assay Buffer: 25 mM Tris, 1 mM CaCl2, 0.5% (w/v) Brij-35, pH 9.0
  • Recombinant Mouse Furin (rmFurin) (Catalog # 6450-SE)
  • Substrate: p-Glu-Arg-Thr-Lys-Arg-AMC (Catalog # ES013), 8 mM in deionized water
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmFurin to 2 μg/mL in Assay Buffer.
  2. Dilute Substrate to 100 µM in Assay Buffer.
  3. Load into a black well plate 50 µL of the 2 μg/mL rmFurin and start the reaction by adding 50 µL of 100 µM substrate.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nM (top read), respectively, in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard 7-amino, 4-Methyl Coumarin (Sigma, Catalog # A-9891).

Per Well:

  • rmFurin: 0.1 µg
  • Substrate: 50 µM

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