Recombinant Human Proprotein Convertase 2/PCSK2 Protein, CF

R&D Systems | Catalog # 6018-SE

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Key Product Details

  • R&D Systems CHO-derived Recombinant Human Proprotein Convertase 2/PCSK2 Protein (6018-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

P16519

Applications

Enzyme Activity
View all Proprotein Convertase 2/PCSK2 Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human Proprotein Convertase 2/PCSK2 protein
Gly110-Asn638 with a C-terminal 6-His tag

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

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

N-terminal Sequence Analysis

Gly110

Predicted Molecular Mass

59 kDa

SDS-PAGE

66-82 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate pERTKR-AMC (Catalog # ES013).
The specific activity is >180 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

6018-SE
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and CaCl2.
Shipping The product is shipped with dry ice or equivalent. 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, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Background: Proprotein Convertase 2/PCSK2

Proprotein Convertase 2 (PCSK2) is a subtilisin-like serine peptidase that processes proteins into biologically active products in the secretory pathway. Like other furin/kexin family members, PCSK2 cleaves its substrates primarily after sites of paired basic amino acid residues (1). Autoactivation of PCSK2 occurs in a post‑Golgi compartment of the secretory system (2). Interaction with the secretory protein 7B2 is required for proper activation of PCSK2 (3). The N-terminal domain of 7B2 stabilizes active PCSK2, and a C-terminal fragment of 7B2 can inhibit PCSK2 activity (4). rhPCSK2 was coexpressed with 7B2 to facilitate activation of the enzyme. PCSK2 is a major proteolytic processing enzyme of the regulated secretory pathway of the neuroendocrine system, where it generates a number of hormones and neuropeptides (5). Products of PCSK2 processing include enkephalins, insulin, somatostatin, dynorphin, and LHRH. 

References

  1. Zhou, A. et al. (1999) J. Biol. Chem. 274:20745.
  2. Lamango, N.S. et al. (1999) Arch. Biochem. Biophys. 362:275.
  3. Zhu, X. and I. Lindberg (1995) J. Cell Biol. 129:1641.
  4. Lamango, N. et al. (1996) Arch. Biochem. Biophys. 330:238.
  5. Rouillé, Y. et al. (1995) Front. Neuroendocrinol. 16:322.

Long Name

Proprotein Convertase Subtilisin/Kexin Type 2

Alternate Names

NEC2, PC2, PCSK2, SPC2

Entrez Gene IDs

5126 (Human); 18549 (Mouse); 25121 (Rat)

Gene Symbol

PCSK2

UniProt

P16519 (Human)

Additional Proprotein Convertase 2/PCSK2 Products

Product Documents for Recombinant Human Proprotein Convertase 2/PCSK2 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 Human Proprotein Convertase 2/PCSK2 Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Human Proprotein Convertase 2/PCSK2 Protein, CF (6018-SE):

Materials
  • Assay Buffer: 50 mM NaOAc, 100 mM NaCl, 1 mM CaCl2, 0.5% (w/v) Brij-35, pH 5.0
  • Recombinant Human Proprotein Convertase 2/PCSK2 (rhPCSK2) (Catalog # 6018-SE)
  • Substrate: L-PYROGlu-Arg-Thr-Lys-Arg-AMC (Catalog # ES013), 4 mM stock in deionized water
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhPCSK2 to 0.4 µg/mL in Assay Buffer.
  2. Dilute Substrate to 500 µM in Assay Buffer.
  3. Start the reaction by mixing equal volumes of 0.4 µg/mL rhPCSK2 and 500 µM Substrate together and incubate at 37 °C for 30 minutes. For a Substrate Blank, mix Assay Buffer and Substrate together and incubate at 37 °C for 30 minutes.
  4. Into a plate, load 100 µL of the reaction and Substrate Blank (in triplicate).
  5. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in endpoint mode.
  6. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Fluorescence* (RFU) x Conversion Factor** (pmol/RFU)
Incubation time (min) x amount of enzyme (µg)

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

Per Well:

  • rhPCSK2: 0.02 µg
  • Substrate: 250 µM

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