As the only known physiological activator of thrombin, Factor X is a vitamin K-dependent plasma protease that plays a pivotal role in blood coagulation. Human Factor X is initially synthesized in the liver as a single-chain precursor of 488 amino acid residues with a signal peptide and a pro region (residues 1‑40). Both the intrinsic and extrinsic pathways activate Factor X to Xa, which consists of light (residues 41‑179) and heavy (residues 235‑488) chains linked by a disulfide bond. The light chain contains a Gla and two EGF-like domains and the heavy chain corresponds to the serine protease domain. The full-length human Factor X was expressed and the pro enzyme was purified and activated. The active protease (rhXa) was further purified and analyzed for its activity towards either peptides or proteins containing a Xa cleavage site. In addition to the activity described in the Activity Assay Protocol, rhFX also be used to cleave fusion proteins containing a Factor Xa cleavage site. The conditions for the optimal cleavage of a particular fusion protein, such as the molar ratio between rhFX and the protein and time and temperature of incubation, are protein-dependent and need to be individually determined.
Recombinant Human Coagulation Factor Xa Protein, CF
R&D Systems | Catalog # 1063-SE
Key Product Details
- R&D Systems Sf 21 (baculovirus)-derived Recombinant Human Coagulation Factor Xa Protein (1063-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
Accession Number
Structure / Form
Applications
Product Specifications
Source
Leu24-Lys488 with a C-terminal 10-His tag
Proform Factor X was expressed, purified, activated and further purified to yield Factor Xa.
Purity
Endotoxin Level
N-terminal Sequence Analysis
Predicted Molecular Mass
SDS-PAGE
Activity
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002).
The specific activity is >700 pmol/min/µg, as measured under the described conditions.
Reviewed Applications
Read 1 review rated 5 using 1063-SE in the following applications:
Formulation, Preparation, and Storage
1063-SE
| Formulation | Lyophilized from a 0.2 μm filtered solution in MES, NaCl and CaCl2. |
| Reconstitution | Reconstitute at 100 μg/mL in sterile 25 mM MES, 150 mM NaCl and 5 mM CaCl2, pH 6.0. |
| 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.
|
Calculators
Background: Coagulation Factor X
Alternate Names
Gene Symbol
UniProt
Additional Coagulation Factor X Products
Product Documents for Recombinant Human Coagulation Factor Xa Protein, CF
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Coagulation Factor Xa Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human Coagulation Factor Xa Protein, CF
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Protocols
View specific protocols for Recombinant Human Coagulation Factor Xa Protein, CF (1063-SE):
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% Brij-35, pH 7.5 (TCNB)
- Recombinant Human
Coagulation Factor Xa (rhFactor Xa) (Catalog # 1063-SE)
- Substrate: Fluorogenic Peptide Substrate II, MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(DNP)-NH2 (Catalog # ES002)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhFactor Xa to 0.4 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load 50 µL of the 0.4 ng/µL rhFactor Xa into plate and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 20 µM Substrate without any rhFactor Xa.
- Read at excitation and emission wavelengths of 320 nm and 405 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 MCA-Pro-Leu-OH (Bachem, Catalog # M-1975)
Per Well:
- rhFactor Xa: 0.02 µg
- Substrate: 10 µM
Associated Pathways
