Coagulation Factor X/F10 蛋白, 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.

凝血因子 X，也称为 F10，是一种维生素 K 依赖性糖蛋白，对于凝血过程至关重要。Xa 因子是 X 因子的活化形式，在存在因子 Va、钙和磷脂的情况下将凝血酶原转化为凝血酶，在凝血级联中发挥关键作用。这种酶促转化是形成稳定血凝块的关键步骤，对于防止过度出血至关重要。X因子与其辅助因子和底物的相互作用强调了它在调节止血的复杂分子事件网络中的重要性。

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

• 
  Greater than 90% as determined by reducing SDS-PAGE.

无菌粉末

Lyophilized from a 0.2 μm filtered solution of 20 mM MES,150 mM NaCl, 0.2 mM CaCl₂, pH 5.5 or pH 6.0.

<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.