Protein C, also known as Coagulation Factor XIV, is a vitamin K-dependent serine protease synthesized in the liver as a single-chain precursor (1). The N‑terminus consists of a signal peptide (amino acid (aa) 1-32) and a propeptide (aa 33-42). The mature chain (aa 43‑461) is converted to two disulfide-linked chains (light: aa 43‑199 and heavy: 200-461) and both forms are inactive. The light chain consists of Gla (gamma-carboxy-glutamate) domain and two EGF-like domains. The heavy chain consists of an activation peptide (aa 200‑211) and a serine protease domain (aa 212-450). Present in plasma at 3 to 5 mg/L, protein C plays a key role in anticoagulation. Physiologically, the inactive forms of protein C are converted to the active form by thrombin, which releases the activation peptide. The active protein C cleaves factor VIIIa and Va to inactivate them. This anticoagulation activity can be enhanced by a presence of a cofactor such as protein S. In hereditary thrombophilia, protein C deficiency is caused by a genetic mutation which affect protein C activity. A severe recessive form may result in a massive thrombosis, which is fatal to the patient. The recombinant human Protein C consists of both the mature chain and the two disulfide-linked chains, which can be activated by treatment with thermolysin.
Recombinant Human Coagulation Factor XIV/Protein C, CF
R&D Systems | Catalog # 3349-SE
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Key Product Details
- R&D Systems NS0-derived Recombinant Human Coagulation Factor XIV/Protein C (3349-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
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived human Coagulation Factor XIV/Protein C protein
Met1-Pro461, with a C-terminal 10-His tag
Met1-Pro461, 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
Ala43 (mature and light chains) & Asp200 (heavy chain)
Predicted Molecular Mass
49 kDa (mature), 31 kDa (heavy), 18 kDa (light)
SDS-PAGE
60-64 kDa, 41-44 kDa and 22 kDa, reducing conditions
Activity
Measured by its ability to cleave the fluorogenic peptide substrate Boc-VPR-AMC (Catalog # ES011).
The specific activity is >25 pmol/min/µg, as measured under the described conditions.
The specific activity is >25 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
3349-SE
| Formulation | Supplied as a 0.2 μm filtered solution in Sodium Acetate and NaCl. |
| Reconstitution |
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|
| 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.
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Background: Coagulation Factor XIV/Protein C
References
- Shen, L. and B. Dahlbäck (2004) in Handbook of Proteolytic Enzymes, Barrett, A.J. et al. eds. p. 1673.
Long Name
Vitamin K-dependent Protein C
Alternate Names
Autoprothrombin IIA, PROC, Protein C
Gene Symbol
PROC
UniProt
Additional Coagulation Factor XIV/Protein C Products
Product Documents for Recombinant Human Coagulation Factor XIV/Protein C, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Coagulation Factor XIV/Protein C, CF
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Human Coagulation Factor XIV/Protein C, CF (3349-SE):
Materials
- Activation Buffer: 50 mM Tris, 150 mM NaCl, 10 mM CaCl2, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
- Assay Buffer: 50 mM Tris, 100 mM NaCl, 0.01% (w/v) Brij-35, pH 8.5
- Recombinant Human Coagulation Factor XIV/Protein C (rhPROC) (Catalog # 3349-SE)
- Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
- 1,10-Phenanthroline (Sigma, Catalog # 320056), 0.6 M stock in DMSO
- Fluorogenic Peptide Substrate: BOC-Val-Pro-Arg-AMC (Catalog # ES011)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhPROC to 100 µg/mL in Activation Buffer with 2.3 µg/mL Thermolysin.
- Incubate for 30 minutes at 37 °C.
- Stop Thermolysin activity by adding 1,10-Phenanthroline to 4 mM.
- Incubate for 15 minutes at room temperature.
- Dilute rhPROC to 4 ng/µL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- Load into a black well plate 50 µL of 4 ng/µL rhPROC, 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 200 µM Substrate.
- Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
- 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 (AMC) (Sigma, Catalog # A-9891).
Per Well:
- rhPROC: 0.200 µg
- Substrate: 100 μM
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Associated Pathways
