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Active Coagulation Factor XII (F12)

FXII; HAF; Contact Factor; Hageman Factor

ACTIVITY TEST

Coagulation factor XII (FXII), a S1A serine protease, is mainly produced in the liver and circulates in plasma as a single chain zymogen. F12 plays a role in initiating the contact activation pathway during clotting. When blood comes into contact with a foreign body, F12 is activated, initiating a series of reactions that eventually lead to the production of thrombin and the formation of fibrin, which promotes blood clotting. In addition, the combination of Plasminogen (Plg) with F12 can promote plasminogen activation and thus increase plasminogen production, which plays an important role in thrombolysis and tissue repair. Thus a functional binding ELISA assay was conducted to detect the interaction of recombinant mouse F12 and recombinant human Plg. Briefly, biotin-linked F12 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 ul were then transferred to Plg-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with Streptavidin-HRP for 30min, then wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µl stop solution to the wells and read at 450 nm immediately. The binding activity of recombinant mouse F12 and recombinant human Plg was shown in Figure 1, the EC50 for this effect is 18.34 ug/mL.

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.

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Magazine Citations
Perfusion Ex vivo simulation of cardiopulmonary bypass with human blood for hemocompatibility testing PubMed: 26243277
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