Serpin F2 is a member of the Serpin superfamily and the primary physiological inhibitor of the serine protease plasmin, which is responsible for the dissolution of fibrin clots (1, 2). In addition to plasmin, Serpin F2 is also an efficient inhibitor of trypsin and chymotrypsin (3). Liver and kidney are major sites of Serpin F2 production and other tissues such as muscle, intestine, central nervous system, and placenta also express its mRNA at a moderate level. The tissue expression pattern of Serpin F2 indicates that it is a key regulator of plasmin-mediated proteolysis in these tissues (4). Human Serpin F2 is synthesized as a 491 amino acid precursor with a 27 amino acid signal peptide. The secreted protein has a short propeptide (residues 28-39) and a mature chain (residues 40-491). The presence of the propeptide did not affect its ability to inhibit plasmin but reduced its cross-linking ability to fibrin (5).
Recombinant Human Serpin F2/alpha 2-Antiplasmin Protein, CF
R&D Systems | Catalog # 1470-PI
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Key Product Details
- R&D Systems NS0-derived Recombinant Human Serpin F2/alpha 2-Antiplasmin Protein (1470-PI)
- 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
Inhibition Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived human Serpin F2/alpha 2-Antiplasmin protein
Met28-Lys491, with a C-terminal 10-His tag
Met28-Lys491, with a C-terminal 10-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
Met28
Predicted Molecular Mass
53 kDa
SDS-PAGE
67 kDa, reducing conditions
Activity
Measured by its ability to inhibit trypsin cleavage of a fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002).
The IC50 value is <2.0 nM, as measured under the described conditions.
The IC50 value is <2.0 nM, as measured under the described conditions.
Formulation, Preparation, and Storage
1470-PI
| Formulation | Lyophilized from a 0.2 μm filtered solution in Tris and NaCl. |
| Reconstitution | Reconstitute at 100 μg/mL in sterile 25 mM Tris and 150 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: Serpin F2/alpha 2-Antiplasmin
References
- Tone, M. et al. (1987) J. Biochem. 102:1033.
- Silverman, G.A. et al. (2001) J. Biol. Chem. 276:33293.
- Potempa, J. et al. (1988) Science 241:699.
- Menoud, P.-A. et al. (1996) J. Clin. Invest. 97:2478.
- Sumi, Y. et al. (1989) J. Biochem. 106:703.
Alternate Names
alpha 2-Antiplasmin
Gene Symbol
SERPINF2
UniProt
Additional Serpin F2/alpha 2-Antiplasmin Products
Product Documents for Recombinant Human Serpin F2/alpha 2-Antiplasmin 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 Serpin F2/alpha 2-Antiplasmin Protein, CF
For research use only
Citations for Recombinant Human Serpin F2/alpha 2-Antiplasmin Protein, CF
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Protocols
View specific protocols for Recombinant Human Serpin F2/alpha 2-Antiplasmin Protein, CF (1470-PI):
Materials
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
- Recombinant Human Serpin F2/ alpha 2‑Antiplasmin (rhSerpin F2 (Catalog # 1470-PI)
- Trypsin (Sigma, Catalog # T-1426)
- 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
- Dilute Trypsin to 0.25 µg/mL in Assay Buffer and KEEP ON ICE.
- Prepare a curve of rhSerpin F2 (MW: 53,056 Da) in Assay Buffer. Make the following serial dilutions: 600, 200, 100, 50, 30, 20, 15, 10, 5 and 1 nM.
- Combine equal volumes of 0.25 µg/mL Trypsin and rhSerpin F2 serial dilutions. Include two enzyme controls of equal volumes of Assay Buffer and 0.25 µg/mL Trypsin.
- Incubate reaction mixtures at 37 °C for 15 minutes.
- Dilute incubated reaction mixtures by 1/5 in Assay Buffer.
- Dilute Substrate to 20 µM with Assay Buffer.
- In a plate load 50 µL of the diluted reaction mixtures, and start the reaction by adding 50 µL of 20 µM Substrate to wells.
- Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
- Derive the 50% inhibition concentration (IC50) for rhSerpin F2 by plotting RFU/min (or specific activity) vs. concentration with 4-PL fitting.
- The specific activity for Trypsin 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 MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).
Per Well:
- Trypsin: 0.00125 µg
- rhSerpin F2 curve: 30, 10, 5, 2.5, 1.5, 1, 0.75, 0.5, 0.25 and 0.05 nM
- Substrate: 10 µM
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Associated Pathways
