Coagulation factor VII/F7 Protein, Human (406a.a, HEK293, His)

Coagulation factor VII (F7) initiates the extrinsic pathway of blood coagulation. As a circulating zymogen, F7 is activated to F7a through proteolysis induced by factor Xa, factor XIIa, factor IXa, or thrombin. In the presence of tissue factor and calcium ions, F7a catalyzes limited proteolysis of factor X, converting it to factor Xa. F7a can also convert factor IX to factor IXa, emphasizing its pivotal role in coagulation cascades. Coagulation factor VII/F7 Protein, Human (406a.a, HEK293, His) is the recombinant human-derived Coagulation factor VII/F7, expressed by HEK293, with C-His labeled tag. The total length of Coagulation factor VII/F7 Protein, Human (406a.a, HEK293, His) is 406 a.a..

Coagulation factor VII (F7) plays a crucial role in initiating the extrinsic pathway of blood coagulation. As a serine protease circulating in the blood in its zymogen form, factor VII undergoes activation to factor VIIa through minor proteolysis induced by factor Xa, factor XIIa, factor IXa, or thrombin. In the presence of tissue factor and calcium ions, factor VIIa further catalyzes the limited proteolysis of factor X, converting it to factor Xa. Additionally, in the presence of tissue factor and calcium, factor VIIa is capable of converting factor IX to factor IXa, highlighting its pivotal function in the intricate cascade of coagulation processes.

Measured by its ability to cleave 100μM fluorogenic peptide substrate Boc-VPR-AMC (HY-P137784) in kinetic mode for 5 minutes. The specific activity is ≥105.77 pmol/min/µg. (Activation description: The proenzyme needs to be activated by Tissue Factor (HY-P7787) for an activated form.)

Materials
Assay buffer: 50 mM Tris, 2.5 mM CaCl₂, pH 8.5
Coagulation factor VII/F7 Protein, Human (406a.a, HEK293, His) (HY-P78763)
Coagulation factor III/F3 Protein, Human (HEK293, His) (Tissue Factor, HY-P7787)
Substrate: Boc-Val-Pro-Arg-AMC hydrochloride (HY-137784)
Standard: 7-amino, 4-Methyl Coumarin (HY-D0027)

Procedure
1. Dilute 7-amino, 4-Methyl Coumarin to 0, 9.765625, 19.53125, 39.0625, 78.125, 156.25, 312.5, 625, 1250, 2500, 5000, 10000 pmol with assay buffer respectively.
2. Add 100 μL of each concentration of standard to the wells.
3. Read in kinetic mode for 5 minutes with excitation and emission wavelengths of 380 nm and 460 nm (top read).
4. Plot a standard curve with the standard concentration as the abscissa and the measured RFU as the ordinate to obtain the standard curve.
5. Protein activity determination: First reconstitute Coagulation factor VII with sterile water to 100 μg/mL, then dilute Coagulation factor VII to 32 μg/mL with assay buffer.
6. Dilute Tissue Factor to 35.5 μg/mL with assay buffer.
7. Mix equal volumes of Coagulation factor VII and Tissue Factor, and incubate at 37°C for 5 minutes.
8. Dilute the substrate to 200 μM in assay buffer.
9. Transfer 50 μL of activated Coagulation factor VII to the wells of a black well plate.
10. Add 50 μL of 200 μM substrate to each well to start the reaction.
11. Read for 5 minutes in kinetic mode at excitation and emission wavelengths of 380 nm and 460 nm respectively.
12. Calculate the specific activity:

Human

HEK293

C-6*His

P08709-1 (A61-P466)

2155  [NCBI]

Full Length of Isoform-1 Mature Protein

ANAFLEELRPGSLERECKEEQCSFEEAREIFKDAERTKLFWISYSDGDQCASSPCQNGGSCKDQLQSYICFCLPAFEGRNCETHKDDQLICVNENGGCEQYCSDHTGTKRSCRCHEGYSLLADGVSCTPTVEYPCGKIPILEKRNASKPQGRIVGGKVCPKGECPWQVLLLVNGAQLCGGTLINTIWVVSAAHCFDKIKNWRNLIAVLGEHDLSEHDGDEQSRRVAQVIIPSTYVPGTTNHDIALLRLHQPVVLTDHVVPLCLPERTFSERTLAFVRFSLVSGWGQLLDRGATALELMVLNVPRLMTQDCLQQSRKVGDSPNITEYMFCAGYSDGSKDSCKGDSGGPHATHYRGTWYLTGIVSWGQGCATVGHFGVYTRVSQYIEWLQKLMRSEPRPGVLLRAPFP

approximately 55-65 kDa

• 
  Greater than 95% as determined by reducing SDS-PAGE.

Lyophilized powder

Lyophilized from a 0.22 μm filtered solution of 50 mM Tris-HCl, 150 mM NaCl, pH 7.5, 8% trehalose.
Note: For SPR assay, please replace the buffer. Primary amine components (e.g., Tris, imidazole) can affect protein-coupled chips.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O.

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.