Recombinant Mouse Coagulation Factor VII Protein, CF

Catalog # Availability Size / Price Qty
3305-SE-010
Product Details
Citations (1)
FAQs
Supplemental Products
Reviews

Recombinant Mouse Coagulation Factor VII Protein, CF Summary

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.
Activity
Measured by its ability to cleave the fluorogenic peptide substrate Boc-VPR-AMC (Catalog # ES011). The specific activity, is >4 pmol/min/µg, as measured under the described conditions.
Source
Chinese Hamster Ovary cell line, CHO-derived mouse Coagulation Factor VII protein
Ala42-Leu446, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
Ala42
Structure / Form
Mature form
Predicted Molecular Mass
47 kDa
SDS-PAGE
58 kDa and 54 kDa, reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

3305-SE

Formulation Lyophilized from a 0.2 μm filtered solution in Tris and NaCl.
Reconstitution Reconstitute at 200 μg/mL in sterile 50 mM Tris, 10 mM CaCl2, 150 mM NaCl and 0.05% Brij-35 (pH 7.5).
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.

Assay Procedure

Materials
  • Activation Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
  • Assay Buffer: 50 mM Tris, pH 9.0
  • Recombinant Mouse Coagulation Factor VII (rmFactor VII) (Catalog # 3305-SE)
  • Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
  • 1,10-Phenanthroline (Sigma, Catalog # 320056)
  • Recombinant Mouse Coagulation Factor III/Tissue Factor (rmTF) (Catalog # 3178-PA)
  • 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. Activate rmFactor VII at 100 µg/mL with 10 µg/mL Thermolysin in Activation Buffer.
  2. Incubate at 37 °C for 30 minutes.
  3. After incubation, stop reaction with 1,10-Phenanthroline at a final concentration of 10 mM in Activation Buffer.
  4. Incubate reaction mixtures at 37 °C for 5 minutes.  The enzyme concentration is now at 75 µg/mL.
  5. Dilute rmTF to 15.3 µg/mL in Assay Buffer.
  6. In a plate, load 13.3 µL of 75 µg/mL activated rmFactor VII followed by adding 36.7 µL of 15.3 µg/mL rmTF.
  7. Incubate plate at 37 °C for 5 minutes.
  8. Dilute Substrate to 200 µM in Assay Buffer.
  9. After plate incubation, start the reaction by adding 50 µL of 200 µM of Substrate to wells.
  10. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively in kinetic mode for 5 minutes.
  11. 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:
  • rmFactor VII: 1 µg
  • rmTF: 0.56 µg
  • Substrate: 100 µM
Reconstitution Calculator

Reconstitution Calculator

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: Coagulation Factor VII

Coagulation Factors VII and VIIa refer to the pro and active forms of the same protease, respectively (1). Factor VII is synthesized in the liver and circulates in the plasma where it binds to tissue factor (TF), an integral membrane protein found in a variety of cell types. Upon binding of TF, factor VII is rapidly converted into 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). The cleavage and activation of Coagulation Factors VII, IX and X by VIIa:TF is phospholipid-dependent whereas the cleavage of small peptide substrates is not (1). The deduced amino acid sequence of mouse factor VII predicts a signal peptide (residues 1 to 24), propeptide (residues 25 to 41), and the mature chain that can be further processed into the light chain (residues 42 to 193) and the heavy chain (residues 194 to 446). The purified recombinant mouse F7 corresponds to the mature chain, which can be processed and activated by treatment with thermolysin and binding with recombinant mouse Tissue Factor (Catalog # 3178-PA) under the conditions described in the Activity Assay Protocol.

References
  1. Morrissey, J.H. (2004) in Handbook of Proteolytic Enzymes, Barrett, A.J. et al. (eds. ), Academic Press, San Diego, p. 1659.
  2. Versteeg, H.H. et al. (2003) Carcinogenesis 24:1009.
Long Name
Coagulation Factor VII (Serum Prothrombin Conversion Accelerator)
Entrez Gene IDs
2155 (Human); 14068 (Mouse)
Alternate Names
coagulation factor VII (serum prothrombin conversion accelerator); Coagulation Factor VII; EC 3.4.21; EC 3.4.21.21; Eptacog alfa; F7; FVII coagulation protein; proconvertin; Serum prothrombin conversion accelerator; SPCA

Citation for Recombinant Mouse Coagulation Factor VII Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Targeting tumor endothelial marker 8 in the tumor vasculature of colorectal carcinomas in mice.
    Authors: Fernando S, Fletcher BS
    Cancer Res., 2009;69(12):5126-32.
    Species: Mouse
    Sample Types: Buffer
    Applications: Enzyme Assay

FAQs

No product specific FAQs exist for this product, however you may

View all Proteins and Enzyme FAQs

Reviews for Recombinant Mouse Coagulation Factor VII Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Mouse Coagulation Factor VII Protein, CF and earn rewards!

Have you used Recombinant Mouse Coagulation Factor VII Protein, CF?

Submit a review and receive an Amazon gift card.

$25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image

$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image

Submit a Review