Active Tissue Factor Pathway Inhibitor (TFPI)

TFI; EPI; LACI; TFPI1; Lipoprotein-Associated Coagulation Inhibitor; Extrinsic pathway inhibitor

ACTIVITY TEST

TFPI, also known as lipoprotein-associated coagulation inhibitor (LACI) and extrinsic pathway inhibitor (EPI), is a physiological inhibitor of extrinsic pathway of coagulation and has biological functions of anticoagulation and anti-inflammation. It is a secreted protein with a N-terminal acidic region, three Kunitz (K) domains separated with by two linker regions, and a C-terminal basic region. The activity of recombinant rat TFPI was measured by its ability to inhibit trypsin cleavage of a fluorogenic peptide substrate Mca-RPKPVE-Nval-WRK(Dnp)-NH2 in the assay buffer 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5. Trypsin was diluted to 50 ug/ml in the assay buffer and 20 ul different concentrations of recombinant rat TFPI (MW: 61.9 KD) was incubated with 20 ul diluted trypsin at 37 ℃ for 15 minutes. Loading 50 µL of the incubated mixtures which were diluted five-fold in assay buffer into empty wells of a plate, and start the reaction by adding 50 µL of 20 µM substrate. Include a substrate blank containing 50 µL of assay buffer and 50 µL of 20 µM substrate. Then read at excitiation and emission wavelengths of 320 nm and 405 nm, respectively, in kinetic mode for 5 minutes. The result was shown in Figure 1 and it was obvious that recombinant rat TFPI significantly decreased trypsin activity. The inhibition IC50 was <10 nM.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

GIVEAWAYS

INCREMENT SERVICES

Magazine Citations
South African Journal of Child Health Tissue factor pathway inhibitor in paediatric patients with nephrotic syndrome Sajch: 383
the journal of immunology Cathepsin G in Experimental Tuberculosis: Relevance for Antibacterial Protection and Potential for Immunotherapy PubMed: 26320257
Molecular Nutrition & Food Research Trans‐fatty acid promotes thrombus formation in mice by aggravating antithrombogenic endothelial functions via Toll‐like receptors PubMed: 25546502
Molecular Carcinogenesis Endothelial cell‐anchored tissue factor pathway inhibitor regulates tumor metastasis to the lung in mice PubMed: 25945811
Cell Biology International Extracellular vesicles from malignant effusions induce tumor cell migration: inhibitory effect ofLMWH tinzaparin. pubmed:27435911
International Journal of Infectious Diseases Kunitz-type protease inhibitor as a vaccine candidate against schistosomiasis mansoni pubmed:29128645
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