Recombinant Human Proprotein Convertase 1/PCSK1 Protein, CF

Catalog # Availability Size / Price Qty
2810-SE-010
Product Details
Citations (2)
FAQs
Supplemental Products
Reviews

Recombinant Human Proprotein Convertase 1/PCSK1 Protein, CF Summary

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.
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.
Source
Mouse myeloma cell line, NS0-derived human Proprotein Convertase 1/PCSK1 protein
Ser111-Arg617, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
Ser111
Structure / Form
Mature form
Predicted Molecular Mass
57 kDa
SDS-PAGE
63-80 kDa, reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

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.

Assay Procedure

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
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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.
Entrez Gene IDs
5122 (Human)
Alternate Names
NEC1; PCSK1; Proprotein Convertase 1

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

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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  1. Ferric ions inhibit proteolytic processing of progastrin.
    Authors: Bramante G, Patel O, Shulkes A, Baldwin GS
    Biochem. Biophys. Res. Commun., 2011;404(4):1083-7.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay
  2. A proprotein convertase-inhibiting serpin with an endoplasmic reticulum targeting signal from Branchiostoma lanceolatum, a close relative of vertebrates.
    Authors: Bentele C, Kruger O, Todtmann U, Oley M, Ragg H
    Biochem. J., 2006;395(3):449-56.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay

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Fluorogenic Peptide Substrates

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