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Active Carboxypeptidase B2 (CPB2)

TAFI; CPU; PCPB; Thrombin Activatable Fibrinolysis Inhibitor; Plasma Carboxypeptidase B; Carboxypeptidase U

ACTIVITY TEST

Carboxypeptidase B2 (CPB2), also known as carboxypeptidase U (CPU) and thrombin-activatable fibrinolysis inhibitor (TAFI), is a member of the peptidase M14 family.
CPB2 is synthesized by the liver and circulates in plasma as a plasminogen-bound zymogen. It can cleave C-terminal arginine or lysine residues from biologically active peptides such as kinins or anaphylatoxins in the circulation thereby regulating their activities and Down-regulates fibrinolysis by removing C-terminal lysine residues from fibrin that has already been partially degraded by plasmin. Besides, Plasminogen (Plg) has been identified as an interactor of CPB2, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human CPB2 and recombinant mouse Plg. Briefly, CPB2 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to Plg-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-CPB2 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, 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 450/630 nm immediately. The binding activity of recombinant human CPB2 and recombinant mouse Plg was shown in Figure 1, the EC50 for this effect is 2.9 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
Taiwan J Obstet Gynecol.  Levels of thrombin-activatable fibrinolysis inhibitor and platelet-activating factor in recurrent pregnancy loss patients Pubmed:26927250
Experiental And Therapeutic Medicine Plasma proteins as potential targets of abnormal Savda syndrome in asthma patients treated with unique Uighur prescription pubmed:28672924
Clinics and Research in Hepatology and Gastroenterology Thrombin activatable fibrinolysis inhibitor as an indicator of the severity of acute pancreatitis Pubmed:22230218
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