Recombinant Human Serpin C1/Antithrombin-III Protein, CF

R&D Systems | Catalog # 1267-PI

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems NS0-derived Recombinant Human Serpin C1/Antithrombin-III Protein (1267-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

P01008

Applications

Inhibition Activity
View all Serpin C1/Antithrombin-III Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Serpin C1/Antithrombin-III protein
His33-Lys464, with a C-terminal 10-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

His33

Predicted Molecular Mass

50 kDa

SDS-PAGE

55-65 kDa, reducing conditions

Activity

Measured by its ability to inhibit Recombinant Human Coagulation Factor II/Thrombin (Catalog # 1473-SE) cleavage of a fluorogenic peptide substrate Boc-VPR-AMC (Catalog # ES011).
The IC50 value is <5 nM, as measured under the described conditions.

Formulation, Preparation, and Storage

1267-PI
Formulation Lyophilized from a 0.2 μm filtered solution in MES and NaCl.
Reconstitution Reconstitute at 100 μg/mL in sterile 25 mM MES, 150 mM NaCl, pH 6.5.
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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

=
÷

Background: Serpin C1/Antithrombin-III

Serpin C1 is a member of the Serpin superfamily of the serine protease inhibitors (1). It is the principal plasma Serpin of blood clotting proteases and inhibits thrombin as well as several factors such as Xa (2). Similar to Serpins A5 and D1, its thrombin inhibitory activity is enhanced by heparin. Hereditary and acquired Serpin C1 deficiency is the cause of an increased thrombotic tendency in many cases (3). For example, acquired Serpin C1 deficiency is a common condition in sepsis, after major trauma or surgery (4).

References

  1. Silverman, G.A. et al. (2001) J. Biol. Chem. 276:33293.
  2. Chuang, Y.-J. et al. (2001) Biochemistry 40:6670.
  3. Vinazzer, H. (1999) Semin. Thromb. Hemost. 25:257.
  4. Risberg, B. (1998) Blood Coagul. Fibrinolysis Suppl. 3:S3.

Alternate Names

Antithrombin-III, ATIII

Entrez Gene IDs

462 (Human); 11905 (Mouse)

Gene Symbol

SERPINC1

UniProt

P01008

Additional Serpin C1/Antithrombin-III Products

Product Documents for Recombinant Human Serpin C1/Antithrombin-III 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.

Download SDS

Product Specific Notices for Recombinant Human Serpin C1/Antithrombin-III Protein, CF

For research use only

Citations for Recombinant Human Serpin C1/Antithrombin-III Protein, CF

Customer Reviews for Recombinant Human Serpin C1/Antithrombin-III Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Human Serpin C1/Antithrombin-III Protein, CF and earn rewards!

Have you used Recombinant Human Serpin C1/Antithrombin-III Protein, CF?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Protocols

View specific protocols for Recombinant Human Serpin C1/Antithrombin-III Protein, CF (1267-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 C1/Antithrombin-III (rhSerpin C1) (Catalog # 1267-PI)
  • Recombinant Human Coagulation Factor II/Thrombin (Catalog # 1473-SE)
  • Heparin (Sigma, Catalog # H3393), 20 mg/mL stock in deionized water
  • 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
  1. Dilute Thrombin to 1 µg/mL with Heparin at 48.6 µg/mL in Assay Buffer.
  2. Prepare a curve of rhSerpin C1 (MW: 50,378 Da) in Assay Buffer. Make the following serial dilutions: 1000, 500, 250, 125, 62.5, 41.7, 27.8, 13.9, 6.94, and 2.31 nM.
  3. Mix equal volumes of rhSerpin C1 curve dilutions and Thrombin/Heparin mixture. Include a control (in duplicate) containing equal volumes of Assay Buffer and Thrombin/Heparin mixture.
  4. Incubate reaction mixtures at room temperature for 30 minutes.
  5. After incubation, dilute reaction mixtures by 1/5 in Assay Buffer.
  6. Dilute Substrate to 200 µM in Assay Buffer.
  7. In a plate load 50 µL of the diluted reaction mixtures to wells, and start the reaction by adding 50 µL of 200 µM Substrate.
  8. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  9. Derive the 50% inhibition concentration (IC50) for rhSerpin C1 by plotting RFU/min (or specific activity) vs. concentration with 4-PL fitting.
  10. Calculate specific activity for Thrombin at each point 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:

  • Thrombin: 0.005 µg (1.0 nM)
  • rhSerpin C1:  50, 25, 12.5, 6.25, 3.13, 2.09, 1.39, 0.695, 0.347, and 0.116 nM
  • Substrate: 100 µM

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes

Associated Pathways

Blood Coagulation Signaling Pathways Blood Coagulation Signaling Pathway Thumbnail