Recombinant Human Coagulation Factor VII Protein, CF

Catalog # Availability Size / Price Qty
2338-SE-010
Product Details
Citations (2)
FAQs
Supplemental Products
Reviews (1)

Recombinant Human 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 >50 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Coagulation Factor VII protein
Ala39-Pro444, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
Ala39
Structure / Form
Mature form
Predicted Molecular Mass
46 kDa
SDS-PAGE
60 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.

2338-SE

Formulation Supplied as a 0.2 μm filtered solution in Sodium Acetate and NaCl.
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.

Assay Procedure

Materials
  • Processing Buffer: 50 mM Tris, 0.15 M NaCl, 10 mM CaCl2, 0.05% (w/v) Brij-35, pH 7.5
  • Assay Buffer: 50 mM Tris, 2.5 mM CaCl2, pH 8.5
  • Recombinant Human Coagulation Factor VII (rhF7) (Catalog # 2338-SE)
  • Substrate: BOC-Val-Pro-Arg-AMC, (Catalog # ES011), 10 mM stock in DMSO
  • Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
  • 1,10-Phenanthroline (Sigma-Aldrich, Catalog # 320050), 0.6 M in DMSO
  • Recombinant Human Coagulation Factor III/Tissue Factor (rhTF) (Catalog # 2339-PA)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Processing
    1. Incubate rhF7 at 100 µg/mL with Thermolysin at 6.25 µg/mL in Processing Buffer for 20 minutes at 37 °C.
    2. Stop Thermolysin activity by adding 1,10-Phenanthroline, diluted in Processing Buffer, to a final concentration of 10 mM.
    3. Incubation at room temperature for 5 minutes.
  2. Activation
    1. Dilute the above processed rhF7 to 32 µg/mL with Assay Buffer.
    2. Dilute rhTF to 35.5 µg/mL with Assay Buffer.
    3. Combine equal volumes of rhF7 (now at 16 µg/mL) and rhTF (now at 17.75 µg/mL) and incubate for 5 minutes at 37 °C.
  3. Activity Assay
    1. Dilute substrate to 200 µM in Assay Buffer.
    2. Transfer 50 µL of activated rhF7 into wells of a black well plate.
    3. Start the reaction by adding 50 μL of 200 µM substrate per well.
    4. Read (top read) in kinetic mode for 5 minutes at excitation and emission wavelengths of 380 nm and 460 nm, respectively.
    5. Calculate specific activity:

         Specific Activity (pmoles/min/µg) =

    Adjusted Vmax* (RFU/min) x Conversion Factor** (pmole/RFU)
    amount of enzyme (µg)

         *Adjusted for Substrate Blank

         **Derived using calibration standard 7-Amino, 4-Methyl Coumarin (Sigma-Aldrich, Catalog # A9891).

Per Well:
  • rhF7: 0.8 µg
  • rhTF: 0.89 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 predominant splicing variant of Factor VII in normal liver corresponds to the 444 amino acid precursor (3, 4). After a signal peptide (residues 1 to 38), the mature chain can be further processed into the light chain (residues 39 to 190) and the heavy chain (residues 191 to 444). The purified Recombinant Human Factor VII corresponds to the mature chain, which can be processed and activated by treatment with thermolysin and binding with Recombinant Human Coagulation Factor III/Tissue Factor (Catalog # 2339-PA) under the conditions described above.

References
  1. Morrissey, J.H. (2004) in Handbook of Proteolytic Enzymes, Barrett, A.J. et al. eds. p. 1659. 
  2. Versteeg, H.H. et al. (2003) Carcinogenesis 24:1009.
  3. Hagen, F.S, et al. (1986) Proc. Natl. Acad. Sci. USA 83:2412.
  4. O’Hara, P.J. et al. (1987) Proc. Natl. Acad. Sci. USA 84:5158.
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

Citations for Recombinant Human 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.

2 Citations: Showing 1 - 2
Filter your results:

Filter by:

  1. Transcription Factor NFAT5 Promotes Migration and Invasion of Rheumatoid Synoviocytes via Coagulation Factor III and CCL2
    Authors: S Lee, JS Kong, S You, HM Kwon, SA Yoo, CS Cho, WU Kim
    J. Immunol., 2018;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  2. AAV-mediated gene transfer in the perinatal period results in expression of FVII at levels that protect against fatal spontaneous hemorrhage.
    Authors: Binny C, McIntosh J, Della Peruta M, Kymalainen H, Tuddenham EG, Buckley SM, Waddington SN, McVey JH, Spence Y, Morton CL, Thrasher AJ, Gray JT, Castellino FJ, Tarantal AF, Davidoff AM, Nathwani AC
    Blood, 2012;119(4):957-66.
    Species: Human
    Sample Types: Plasma
    Applications: ELISA (Capture)

FAQs

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

View all Proteins and Enzyme FAQs

Reviews for Recombinant Human Coagulation Factor VII Protein, CF

Average Rating: 5 (Based on 1 Review)

5 Star
100%
4 Star
0%
3 Star
0%
2 Star
0%
1 Star
0%

Have you used Recombinant Human 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

Filter by:


Recombinant Human Coagulation Factor VII Protein, CF
By Anonymous on 08/03/2016
Application: Enzymatic activity in vitro