Coagulation Factor II, commonly known as thrombin, is an essential component of the coagulation cascade in which it converts fibrinogen to fibrin, activates factors V, VII, VIII, XIII and forms complexes with protein C and thrombomodulin (1). It also activates platelets and regulates the behavior of additional cells through protease‑activated receptors (PARs) (2). It may have either protective or deleterious functions, depending on the level and location (3). Its activity is regulated by endogenous inhibitors such as anti-thrombin III (serpin C1) or heparin cofactor II (serpin D1). A plasma serine protease, thrombin is synthesized in the liver as a 622 amino acid precursor with a 24 amino acid signal peptide. Cleavage by itself or by similar enzymes converts the proenzyme to three forms designated as alpha ‑, beta ‑ and gamma ‑thrombin. Composed of a disulfide bond-linked dimer of the light chain (A) (residues 328-363) and the heavy chain (B) (residues 364‑622), alpha -thrombin displays the diverse functions as described above. In comparison, the further processed B chains of beta - and gamma -thrombin have no known physiological function, but retain most of the activity towards small synthetic substrates (4). The recombinant human thrombin may be used to process proteins containing the thrombin cleavage site. The conditions for processing different proteins, such as the ratio of protein/thrombin, buffer components, temperature and time of incubation, may vary and should be empirically determined.
Recombinant Human Coagulation Factor II/Thrombin Protein, CF
R&D Systems | Catalog # 1473-SE
Key Product Details
- R&D Systems NS0-derived Recombinant Human Coagulation Factor II/Thrombin Protein (1473-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
Met1-Glu622 with a C-terminal 10-His tag
The proenzyme was purified, activated and further purified.
Purity
Endotoxin Level
N-terminal Sequence Analysis
Predicted Molecular Mass
SDS-PAGE
Activity
The specific activity is >20,000 pmol/min/µg, as measured under the described conditions.
Reviewed Applications
Read 1 review rated 5 using 1473-SE in the following applications:
Formulation, Preparation, and Storage
1473-SE
| Formulation | Lyophilized from a 0.2 μm filtered solution in HEPES and NaCl. |
| Reconstitution | Reconstitute at 100 μg/mL in sterile 50 mM HEPES and 580 mM NaCl, pH 7.5. |
| 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 II/Thrombin
References
- Degen, S.J. and E.W. Davie (1987) Biochemistry 26:6165.
- Coughlin, S.R. (2000) Nature 407:258.
- Xi, G. et al. (2003) J. Neurochem. 84:3.
- Rydel, T.J. et al. (1994) J. Biol. Chem. 269:22000.
Alternate Names
Gene Symbol
UniProt
Additional Coagulation Factor II/Thrombin Products
Product Documents for Recombinant Human Coagulation Factor II/Thrombin 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 Human Coagulation Factor II/Thrombin Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human Coagulation Factor II/Thrombin Protein, CF
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Application: growth of mamospheres.Verified Customer | Posted 03/30/2019Used thrombin for the growth of mamospheres.
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Protocols
View specific protocols for Recombinant Human Coagulation Factor II/Thrombin Protein, CF (1473-SE):
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
- Recombinant Human Coagulation Factor II/Thrombin (rhThrombin) (Catalog # 1473-SE)
- 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 rhThrombin to 0.04 µg/mL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- Load 50 µL of 0.04 µg/mL rhThrombin into a plate, 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 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 (Sigma, Catalog # A-9891).
Per Well:
- rhThrombin: 0.002 µg
- Substrate: 100 µM
FAQs for Recombinant Human Coagulation Factor II/Thrombin Protein, CF
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Q: The product page for Recombinant Human Coagulation Factor II/Thrombin Protein, CF (Catalog # 1473-SE) lists three different molecular weights: light chain A, heavy chain B, and C-terminal fragment. What is the estimated molecular weight of this protein?
A: The active form of thrombin consists of the light alpha chain and the heavy beta chain. With this understanding, the molecular mass of our 1473-SE protein is ~ 35 kDa. The C-terminal fragment, 14 kDa, is a by-product of the activation step and represents a small fraction of the preparation. It has not been removed by any purification steps and appears on SDS-PAGE gels which is the reason we report it.
-
Q: What is the C-terminal fragment mentioned in the Product Description?
A: The 14 kDa C-terminal fragment is a by-product of the activation step and represents a small fraction of the preparation. It has not been removed by any purification steps and appears on SDS-PAGE gels and is the reason we report it.
-
Q: What is the theoretical molecular mass of Recombinant Human Coagulation Factor II/Thrombin Protein (catalog 1473-SE)?
A: The active form of thrombin consists of the light alpha chain and the heavy beta chain. With this understanding, the theoretical molecular mass of our product is ~35 kDa.
-
Q: The product page for Recombinant Human Coagulation Factor II/Thrombin Protein, CF (Catalog # 1473-SE) lists three different molecular weights: light chain A, heavy chain B, and C-terminal fragment. What is the estimated molecular weight of this protein?
A: The active form of thrombin consists of the light alpha chain and the heavy beta chain. With this understanding, the molecular mass of our 1473-SE protein is ~ 35 kDa. The C-terminal fragment, 14 kDa, is a by-product of the activation step and represents a small fraction of the preparation. It has not been removed by any purification steps and appears on SDS-PAGE gels which is the reason we report it.
-
Q: What is the C-terminal fragment mentioned in the Product Description?
A: The 14 kDa C-terminal fragment is a by-product of the activation step and represents a small fraction of the preparation. It has not been removed by any purification steps and appears on SDS-PAGE gels and is the reason we report it.
-
Q: What is the theoretical molecular mass of Recombinant Human Coagulation Factor II/Thrombin Protein (catalog 1473-SE)?
A: The active form of thrombin consists of the light alpha chain and the heavy beta chain. With this understanding, the theoretical molecular mass of our product is ~35 kDa.
-
Q: The product page for Recombinant Human Coagulation Factor II/Thrombin Protein, CF (Catalog # 1473-SE) lists three different molecular weights: light chain A, heavy chain B, and C-terminal fragment. What is the estimated molecular weight of this protein?
A: The active form of thrombin consists of the light alpha chain and the heavy beta chain. With this understanding, the molecular mass of our 1473-SE protein is ~ 35 kDa. The C-terminal fragment, 14 kDa, is a by-product of the activation step and represents a small fraction of the preparation. It has not been removed by any purification steps and appears on SDS-PAGE gels which is the reason we report it.
-
Q: What is the C-terminal fragment mentioned in the Product Description?
A: The 14 kDa C-terminal fragment is a by-product of the activation step and represents a small fraction of the preparation. It has not been removed by any purification steps and appears on SDS-PAGE gels and is the reason we report it.
-
Q: What is the theoretical molecular mass of Recombinant Human Coagulation Factor II/Thrombin Protein (catalog 1473-SE)?
A: The active form of thrombin consists of the light alpha chain and the heavy beta chain. With this understanding, the theoretical molecular mass of our product is ~35 kDa.
Associated Pathways
