Recombinant Human Proprotein Convertase 1/PCSK1 Protein, CF

R&D Systems | Catalog # 2810-SE

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Proprotein Convertase 1/PCSK1 Protein (2810-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

NS0

Accession Number

P29120

Structure / Form

Mature form

Applications

Enzyme Activity
View all Proprotein Convertase 1/PCSK1 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Proprotein Convertase 1/PCSK1 protein
Ser111-Arg617, with a C-terminal 10-His tag

Purity

>95%, 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

Ser111

Predicted Molecular Mass

57 kDa

SDS-PAGE

63-80 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate pERTKR-AMC (Catalog # ES013).
The specific activity is >30 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

2810-SE
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
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 1/PCSK1

The human PCSK1 gene encodes Proprotein Convertase 1 (PC1), which is also known as PC3 (1, 2). As a serine protease of the furin/PC family, the deduced amino acid sequence of human PC1 consists of a signal peptide (residues 1 to 27), a pro peptide (residue 28 to 110), and a mature chain (residues 111 to 753). Autocatalytic processing results in the removal of the pro peptide as well as a region from the C-terminus, which leads to the production of the most active 66 kDa form (3). The purified rhPC1 consists of residues 111 to 617, is active in the assay described above and most likely corresponds to the 66 kDa form. Found almost exclusively in endocrine and neural cells and stored as the 66 kDa form in dense-core secretory vesicles, PC1 is capable of processing several endocrine and neural prohormones, such as opiomelanocortin, somatostatin, enkephalin, insulin, dynorphin and thyrotropin-releasing hormone (3). Indeed, obesity and impaired prohormone processing have been associated with mutations in the human PCSK1 gene (4).

References

  1. Creemers, J.W. et al. (1992) FEBS Lett. 300:82.
  2. Seidah, N.G. et al. (1992) DNA Cell Biol. 11:283.
  3. Seidah, N.G. and M. Chretien (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.) p. 1861. Academic Press, San Diego.
  4. Jackson, R.S. et al. (1997) Nature Genet. 16:303.

Alternate Names

NEC1, PCSK1

Entrez Gene IDs

5122 (Human)

Gene Symbol

PCSK1

UniProt

P29120

Additional Proprotein Convertase 1/PCSK1 Products

Product Documents for Recombinant Human Proprotein Convertase 1/PCSK1 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 1/PCSK1 Protein, CF

For research use only

Citations for Recombinant Human Proprotein Convertase 1/PCSK1 Protein, CF

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Protocols

View specific protocols for Recombinant Human Proprotein Convertase 1/PCSK1 Protein, CF (2810-SE):

Materials
  • Assay Buffer: 25 mM MES, 5 mM CaCl2, 1% (w/v) Brij-35, pH 6.0.
  • Recombinant Human Proprotein Convertase 1/PCSK1 (rhPCSK1) (Catalog # 2810-SE)
  • Substrate: L-PyroGlu-Arg-Thr-Lys-Arg-AMC (pERTKR-AMC) (Catalog # ES013)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhPCSK1 to 4 µg/mL in Assay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer.
  3. Load 50 µL of 4 µg/mL of rhPCSK1 into a plate, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate.
  4. Incubate at 37 °C for 1 hour.
  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 # A9891).

Per Well:

  • rhPCSK1: 0.200 µg
  • Substrate: 100 µM

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