Coagulation Factor III/Tissue Factor (TF), also known as thromboplastin and CD142, is an integral membrane protein found in a variety of cell types. It functions as a protein cofactor/receptor of Coagulation Factor VII, which is synthesized in the liver and circulated in the plasma (1). Upon binding of TF, the inactive factor VII is rapidly converted into activated 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). Synthesized as a 294 amino acid precursor, mouse TF consists of a signal peptide (residues 1 to 28) and the mature chain (residues 29 to 294). As a type I membrane protein, it contains a transmembrane region (residues 252 to 274) and a cytoplasmic tail (residues 275 to 294) (3, 4). The purified rmTF corresponds to the ectodomain (residues 29 to 251) and is potent in activating thermolysin-processed Recombinant Mouse Coagulation Factor VII (Catalog # http://www.rndsystems.com/product_results.aspx?k=3305-SE">3305-SE) under the conditions described in the Activity Assay Protocol.
Recombinant Mouse Coagulation Factor III/Tissue Factor, CF
R&D Systems | Catalog # 3178-PA
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Key Product Details
- R&D Systems NS0-derived Recombinant Mouse Coagulation Factor III/Tissue Factor (3178-PA)
- 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 mouse Coagulation Factor III/Tissue Factor protein
Ala29-Glu251, with a C-terminal 6-His tag
Ala29-Glu251, with a C-terminal 6-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
Ala29
Predicted Molecular Mass
26 kDa
SDS-PAGE
38 kDa and 44 kDa, reducing conditions
Activity
Measured by its ability to activate Coagulation Factor VII in cleaving a fluorogenic peptide substrate Boc-VPR-AMC (Catalog # ES011).
The AC50 is <5 µg/mL, as measured under the described conditions.
Reviewed Applications
Read 1 review rated 5 using 3178-PA in the following applications:
Formulation, Preparation, and Storage
3178-PA
| Formulation | Lyophilized from a 0.2 μm filtered solution in Tris, NaCl and Brij-35. |
| Reconstitution | Reconstitute at 200 μg/mL in sterile 50 mM Tris, pH 9.0. |
| 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 III/Tissue Factor
References
- Morrissey, J.H. (2004) in Handbook of Proteolytic Enzymes. Barrett, A.J. et al. (ed) Academic Press, San Diego, p. 1659.
- Versteeg, H.H. et al. (2003) Carcinogenesis 24:1009.
- Ranganathan, G. et al. (1991) J. Biol. Chem. 266:496.
- Hartzell, S. (1989) Mol. Cell. Biol. 9:2567.
Alternate Names
CD142, F3, Thromboplastin, Tissue Factor
Gene Symbol
F3
UniProt
Additional Coagulation Factor III/Tissue Factor Products
Product Documents for Recombinant Mouse Coagulation Factor III/Tissue Factor, CF
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Mouse Coagulation Factor III/Tissue Factor, CF
For research use only
Related Research Areas
Citations for Recombinant Mouse Coagulation Factor III/Tissue Factor, CF
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Protocols
View specific protocols for Recombinant Mouse Coagulation Factor III/Tissue Factor, CF (3178-PA):
Materials
- Buffer A: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
- Buffer B: 50 mM Tris, pH 9.0
- Recombinant Mouse Coagulation Factor III/Tissue Factor (rmTF) (Catalog # 3178-PA)
- Recombinant Mouse Coagulation Factor VII (rmF7) (Catalog # 3305-SE)
- Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
- 1,10 Phenanthroline (Sigma, Catalog # 320056), 600 mM Stock in DMSO
- Substrate: t-Butyloxycarbonyl-Val-Pro-Arg-7-amido-4-methylcoumarin (Catalog # ES011)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Activate rmF7 at 100 µg/mL in Buffer A containing 10 µg/mL Thermolysin.
- Incubate for 30 minutes at 37 °C.
- Stop reaction by adding 1,10 phenanthroline to a final concentration of 10 mM in Buffer A.
- Incubate for 5 minutes at 37 °C.
- Prepare a curve of rmTF in Buffer B. Make the following serial dilutions: 90, 45, 22.5, 11.3, 5.62, 2.81, 1.41, and 0.703 µg/mL.
- Dilute Thermolysin-activated rmF7 to 32 µg/mL in Buffer B.
- Load in a plate 25 µL of each of the rmTF curve dilutions from step 5. Include two blank controls containing 25 µL of Buffer B and no rmTF.
- Add 25 µL of 32 µg/mL Thermolysin-activated rmF7 to the wells containing rmTF as well as the control wells.
Incubate the reaction in the microplate for 5 minutes at 37 °C. - Dilute Substrate to 200 µM in Buffer B.
- Add 50 µL of 200 µM Substrate to each well containing rmF7 to start the reaction.
- Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
- Derive the 50% activating concentration (AC50) for rmTF by plotting RFU/min (or specific activity) vs. concentration with 4-PL fitting.
- The specific activity for rmF7 at each point may be determined using the following formula (if needed):
|
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:
- rmF7: 0.8 µg
- rmTF: 22.5, 11.3, 5.63, 2.81, 1.41, 0.703, 0.353, and 0.175 µg/mL
- Substrate: 100 µM
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Associated Pathways
