Coagulation Factor X/F10 Protein, Human (HEK293, Fc)

Factor X (F10) is a vitamin K-dependent glycoprotein that plays a key role in the blood coagulation process as factor Xa. Factor Xa, along with factor Va, calcium, and phospholipids, coordinates the conversion of prothrombin to thrombin, a key step in the coagulation cascade. Coagulation Factor X/F10 Protein, Human (HEK293, Fc) is the recombinant human-derived Coagulation Factor X/F10 protein, expressed by HEK293 , with C-hFc labeled tag.

Coagulation Factor X, also known as F10, is a vitamin K-dependent glycoprotein crucial for the blood clotting process. Factor Xa, an activated form of Factor X, plays a pivotal role in the coagulation cascade by converting prothrombin to thrombin in the presence of factor Va, calcium, and phospholipids. This enzymatic conversion is a key step in the formation of a stable blood clot, essential for preventing excessive bleeding. The interaction of Factor X with its cofactors and substrates underscores its significance in the intricate network of molecular events that regulate hemostasis.

Measured by its ability to cleave 20 µM fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2 that incubate at room temperature in kinetic mode for 5 minutes. The specific activity is 1174.20 pmol/min/µg.

Materials
Assay buffer:50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% Brij-35, pH 7.5 (TCNB)
Coagulation Factor X/F10 Protein, Human (HEK293, Fc) (HY-P7860)
Fluorescent Peptide Substrate II, NFF-3 TFA (HY-P2185A)
Standard:MCA-Pro-Leu-OH

Procedure
1. dilute Human Factor Xa to 0.4 ng/μL with assay buffer.
2. Dilute the substrate to 20μM with assay buffer.
3. 50μL of 0.4 ng/ μL Human Factor Xa was added to the wells and 50μL of 20μM substrate was added to start the reaction. Blank control: 50μL of assay buffer and 50μL of 20μM substrate were added to the wells. Do three sets of wells for each concentration.
4. Read in kinetic mode for 5 minutes at excitation and emission wavelengths of 320 nm and 405 nm, respectively.
5. Preparation of standards: MCA-Pro-Leu-OH dilutions were prepared at concentrations of 10 μM, 5 μM, 2.5 μM, 1.25 μM, 0.625 μM, 0.313 μM, 0.156 μM, 0.078 μM, 0.039 μM, 0 μM, and three pairs of wells were made for each gradient, and the fluorescence values were read under the excitation and emission wavelengths of 320 nm and 405 nm respectively. nm to read the fluorescence values.
6. Calculate the specific activity:

Human

HEK293

C-hFc

P00742 (N32-K488)

2159  [NCBI]

Full Length of Mature Protein

NNILARVTRANSFLEEMKKGHLERECMEETCSYEEAREVFEDSDKTNEFWNKYKDGDQCETSPCQNQGKCKDGLGEYTCTCLEGFEGKNCELFTRKLCSLDNGDCDQFCHEEQNSVVCSCARGYTLADNGKACIPTGPYPCGKQTLERRKRSVAQATSSSGEAPDSITWKPYDAADLDPTENPFDLLDFNQTQPERGDNNLTRIVGGQECKDGECPWQALLINEENEGFCGGTILSEFYILTAAHCLYQAKRFKVRVGDRNTEQEEGGEAVHEVEVVIKHNRFTKETYDFDIAVLRLKTPITFRMNVAPACLPERDWAESTLMTQKTGIVSGFGRTHEKGRQSTRLKMLEVPYVDRNSCKLSSSFIITQNMFCAGYDTKQEDACQGDSGGPHVTRFKDTYFVTGIVSWGEGCARKGKYGIYTKVTAFLKWIDRSMKTRGLPKAKSHAPEVITSSPLK

Approximately 80-130 & 22-25 kDa

• 
  ≥ 90%, as determined by reducing SDS-PAGE.

Lyophilized powder

1.Lyophilized from a 0.22 μm filtered solution of 20 mM MES, 150 mM NaCl, 0.2 mM CaCl₂, pH 5.5.
2.Lyophilized from a 0.22 μm filtered solution of 20 mM MES, 150 mM NaCl, 0.2 mM CaCl₂, pH 6.0.
Please refer to the lot-specific COA for specific buffer information.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

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.