Recombinant Mouse Serpin F2/alpha 2-Antiplasmin Protein, CF

R&D Systems | Catalog # 1239-PI

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Serpin F2/alpha 2-Antiplasmin Protein (1239-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

Q61247

Applications

Inhibition Activity
View all Serpin F2/alpha 2-Antiplasmin Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Serpin F2/alpha 2-Antiplasmin protein
Val28-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

Val28

Predicted Molecular Mass

54 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 <1.0 nM, as measured under the described conditions.

Formulation, Preparation, and Storage

1239-PI
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
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: Serpin F2/alpha 2-Antiplasmin

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 (3). The tissue expression pattern of Serpin F2 indicates that it is a key regulator of plasmin-mediated proteolysis in these tissues. Mouse 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 to 39) and a mature chain (residues 40 to 491). For human Serpin F2, the presence of the propeptide did not affect its ability to inhibit plasmin but reduced its cross-linking ability to fibrin (4).

References

  1. Menoud, P.-A. et al. (1996) J. Clin. Invest. 97:2478.
  2. Silverman, G.A. et al. (2001) J. Biol. Chem. 276:33293.
  3. Potempa, J. et al. (1988) Science. 241:699.
  4. Sumi, Y. et al. (1989) J. Biochem. 106:703.
 

Alternate Names

alpha 2-Antiplasmin

Entrez Gene IDs

5345 (Human); 18816 (Mouse)

Gene Symbol

SERPINF2

UniProt

Q61247

Additional Serpin F2/alpha 2-Antiplasmin Products

Product Documents for Recombinant Mouse Serpin F2/alpha 2-Antiplasmin 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 Serpin F2/alpha 2-Antiplasmin Protein, CF

For research use only

Citations for Recombinant Mouse Serpin F2/alpha 2-Antiplasmin Protein, CF

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Protocols

View specific protocols for Recombinant Mouse Serpin F2/alpha 2-Antiplasmin Protein, CF (1239-PI):

Materials
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB
  • Recombinant Mouse Serpin F2/ alpha 2‑Antiplasmin (rmSerpin F2) (Catalog # 1239-PI)
  • Trypsin (Sigma, Catalog # T-1426)
  • Substrate: Mca-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(DNP)-NH2 (Catalog # ES002
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute Trypsin to 0.25 µg/mL in Assay Buffer and KEEP ON ICE.
  2. Prepare a curve of rmSerpin F2 (MW: 53,503 Da) in Assay Buffer. Make the following serial dilutions: 100, 20, 15, 10, 7, 4, 2, and 1 nM.
  3. Combine equal volumes of diluted Trypsin and rmSerpin F2 curve dilutions. Include two blank controls containing equal volumes of Assay Buffer and diluted Trypsin without any rmSerpin F2.
  4. Incubate mixtures at 37 °C for 15 minutes.
  5. Dilute the reaction mixtures 1/5 with Assay Buffer.
  6. Dilute Substrate to 20 µM with Assay Buffer.
  7. Load in a black well plate 50 µL of the incubated mixtures, and start the reaction by adding 50 µL of 20 µM Substrate.
  8. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  9. Derive the 50% inhibiting concentration (IC50) of rmSerping F2 by plotting RFU/min (or specific activity) vs. concentration with 4-PL fitting.
  10. 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
  • rmSerpin F2 curve: 5, 1, 0.75, 0.5, 0.35, 0.2, 0.1, and 0.05 nM
  • Substrate: 10 µM

FAQs

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Associated Pathways

Blood Coagulation Signaling Pathways Blood Coagulation Signaling Pathway Thumbnail