Coagulation factor VII/F7 蛋白, 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..

凝血因子 VII (F7) 在启动外源性凝血途径中发挥着至关重要的作用。作为一种以其酶原形式在血液中循环的丝氨酸蛋白酶，因子 VII 通过因子 Xa、因子 XIIa、因子 IXa 或凝血酶诱导的轻微蛋白水解而活化为因子 VIIa。在组织因子和钙离子存在的情况下，因子 VIIa 进一步催化因子 X 的有限蛋白水解，将其转化为因子 Xa。此外，在存在组织因子和钙的情况下，因子 VIIa 能够将因子 IX 转化为因子 IXa，突出了其在复杂的凝血过程级联中的关键功能。

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.

<1 EU/μg, determined by LAL method.