Coagulation Factors VII and VIIa refer to the pro and active forms of the same protease, respectively (1). Factor VII is synthesized in the liver and circulates in the plasma where it binds to tissue factor (TF), an integral membrane protein found in a variety of cell types. Upon binding of TF, factor VII is rapidly converted into VIIa. The resulting 1:1 complex of VIIa and TF initiates the coagulation pathway and has also important coagulation-independent functions such as angiognesis (2). The cleavage and activation of Coagulation Factors VII, IX and X by VIIa:TF is phospholipid-dependent whereas the cleavage of small peptide substrates is not (1). The deduced amino acid sequence of mouse factor VII predicts a signal peptide (residues 1 to 24), propeptide (residues 25 to 41), and the mature chain that can be further processed into the light chain (residues 42 to 193) and the heavy chain (residues 194 to 446). The purified recombinant mouse F7 corresponds to the mature chain, which can be processed and activated by treatment with thermolysin and binding with recombinant mouse Tissue Factor (Catalog # http://www.rndsystems.com/product_results.aspx?k=3178-PA">3178-PA) under the conditions described in the Activity Assay Protocol.
Recombinant Mouse Coagulation Factor VII Protein, CF
R&D Systems | Catalog # 3305-SE
Key Product Details
- R&D Systems CHO-derived Recombinant Mouse Coagulation Factor VII Protein (3305-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
Ala42-Leu446, with a C-terminal 10-His tag
Purity
Endotoxin Level
N-terminal Sequence Analysis
Predicted Molecular Mass
SDS-PAGE
Activity
The specific activity, is >4 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
3305-SE
| Formulation | Lyophilized from a 0.2 μm filtered solution in Tris and NaCl. |
| Reconstitution | Reconstitute at 200 μg/mL in sterile 50 mM Tris, 10 mM CaCl2, 150 mM NaCl and 0.05% Brij-35 (pH 7.5). |
| Shipping | The product is shipped at ambient temperature. 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 VII
References
- Morrissey, J.H. (2004) in Handbook of Proteolytic Enzymes, Barrett, A.J. et al. (eds. ), Academic Press, San Diego, p. 1659.
- Versteeg, H.H. et al. (2003) Carcinogenesis 24:1009.
Long Name
Alternate Names
Gene Symbol
UniProt
Additional Coagulation Factor VII Products
Product Documents for Recombinant Mouse Coagulation Factor VII Protein, 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 Mouse Coagulation Factor VII Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Mouse Coagulation Factor VII Protein, CF
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Protocols
View specific protocols for Recombinant Mouse Coagulation Factor VII Protein, CF (3305-SE):
- Activation Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
- Assay Buffer: 50 mM Tris, pH 9.0
- Recombinant Mouse Coagulation Factor VII (rmFactor VII) (Catalog # 3305-SE)
- Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
- 1,10-Phenanthroline (Sigma, Catalog # 320056)
- Recombinant Mouse Coagulation Factor III/Tissue Factor (rmTF) (Catalog # 3178-PA)
- Substrate: Boc-Val-Pro-Arg-AMC (Catalog # ES011), 10 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Activate rmFactor VII at 100 µg/mL with 10 µg/mL Thermolysin in Activation Buffer.
- Incubate at 37 °C for 30 minutes.
- After incubation, stop reaction with 1,10-Phenanthroline at a final concentration of 10 mM in Activation Buffer.
- Incubate reaction mixtures at 37 °C for 5 minutes. The enzyme concentration is now at 75 µg/mL.
- Dilute rmTF to 15.3 µg/mL in Assay Buffer.
- In a plate, load 13.3 µL of 75 µg/mL activated rmFactor VII followed by adding 36.7 µL of 15.3 µg/mL rmTF.
- Incubate plate at 37 °C for 5 minutes.
- Dilute Substrate to 200 µM in Assay Buffer.
- After plate incubation, start the reaction by adding 50 µL of 200 µM of Substrate to wells.
- 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 (Sigma, Catalog # A-9891).
Per Well:
- rmFactor VII: 1 µg
- rmTF: 0.56 µg
- Substrate: 100 µM
FAQs for Recombinant Mouse Coagulation Factor VII Protein, CF
-
Q: Is the addition of Coagulation Factor III/Tissue Factor necessary for the activity of Coagulation Factor VII?
A: Our testing has shown minimal detectable kinetic activity of Coagulation Factor VII following cleavage with Thermolysin in the absence of Tissue Factor.
Upon binding with Tissue Factor (TF), Coagulation Factor VII is rapidly converted into Coagulation Factor VIIa. The resulting 1:1 complex of VIIa and TF initiates the coagulation pathway. This complex also has important coagulation-independent functions such as angiogenesis.
