Recombinant Mouse Coagulation Factor Xa Protein, CF

R&D Systems | Catalog # 6724-SE

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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Mouse Coagulation Factor Xa Protein (6724-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

Sf 21 (baculovirus)

Accession Number

O88947

Structure / Form

Recombinant Mouse Coagulation Factor Xa is prone to proteolytic cleavage at C-terminus. The predominant form of the purified protein lacks the His tag.

Applications

Enzyme Activity
View all Coagulation Factor X Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived mouse Coagulation Factor X protein
Met1-Asn481, with a C-terminal 10-His tag.
Accession # O88947
Proform Factor X was expressed, purified, activated and further purified to yield Factor Xa.

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Endotoxin Level

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

N-terminal Sequence Analysis

Tyr84, Ile232

Predicted Molecular Mass

29 kDa & 10 kDa

SDS-PAGE

30-38 kDa & 13-15 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002).
The specific activity is >300 pmol/min/μg, as measured under the described conditions.

Reviewed Applications

Read 1 review rated 5 using 6724-SE in the following applications:

Formulation, Preparation, and Storage

6724-SE
Formulation Supplied as a 0.2 μm filtered solution in MES, NaCl and CaCl2.
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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: Coagulation Factor X

Factor X is the only known physiological activator of thrombin and plays a critical role in blood coagulation. Factor X is also involved in cell surface responses including cell activation, gene expression and mitogenesis. Factor X is a serine protease that is synthesized in the liver and released into the bloodstream. Mouse Factor X consists of a single‑chain precursor of 481 amino acid residues that contains a signal peptide (residues 1‑20), pro region (residues 21‑40), light chain (residues 41‑180) and heavy chain (residues 184‑481). The light and heavy chains are 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 mouse Factor X was expressed and purified. Upon activation, the active protease (rmFactor Xa) was further purified and analyzed for activity against a peptide substrate. In addition to the activity described in the Activity Assay Protocol, Factor X can also be used to cleave fusion proteins containing a Factor X cleavage site. The conditions for the optimal cleavage of a particular fusion protein must be determined for each individual fusion protein.

References

  1. Brown, M. A. et al. (2004) in Handbook of Proteolytic Enzymes (eds. Barrett, A. et al.), pp. 1662, Academic Press, San Diego.

Alternate Names

Cf10, F10, FX, FXa, Prothrombinase, Stuart Factor

Entrez Gene IDs

2159 (Human); 14058 (Mouse); 29243 (Rat)

Gene Symbol

F10

UniProt

O88947

Additional Coagulation Factor X Products

Product Documents for Recombinant Mouse Coagulation Factor Xa 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 Mouse Coagulation Factor Xa Protein, CF

For research use only

Citations for Recombinant Mouse Coagulation Factor Xa Protein, CF

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Protocols

View specific protocols for Recombinant Mouse Coagulation Factor Xa Protein, CF (6724-SE):

Materials
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
  • Recombinant Mouse Coagulation Factor Xa (rmFactor Xa) (Catalog # 6724-SE)
  • Substrate: MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys-(DNP)-NH2 (Catalog # ES002), 2 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmFactor Xa to 0.8 ng/μL in Assay Buffer.
  2. Dilute Substrate to 20 μM in Assay Buffer.
  3. Load into a plate 50 μL of 0.8 ng/μL rmFactor Xa, and start the reaction by adding 50 μL of 20 μM Substrate. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of 20 μM Substrate.
  4. Read plate at excitation and emission wavelengths of 320 nm and 405 nm, respectively, in kinetic mode for 5 minutes.
  5. 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:

  • rmFactor Xa: 0.040 μg
  • Substrate: 10 μM

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