Reagent kit for the optical determination of alpha-2-antiplasmin activity (Rechrom-Antiplasmin) according to TU 9398-007-05595541-2012. Designed for the quantitative determination of alpha-2-antiplasmin activity in blood plasma using a chromogenic method, both manually and on automatic analyzers.
Alpha-2-antiplasmin is a fibrinolysis inhibitor that rapidly binds to plasmin, irreversibly inhibiting its activity.
Principle of the method: The method for determining alpha-2-antiplasmin activity is based on its ability to neutralize plasmin. The activity of alpha-2-antiplasmin in plasma is determined by adding an excess of plasmin to it. This leads to the inhibition of plasmin, which is proportional to the amount of alpha-2-antiplasmin in the plasma. The remaining amount of plasmin catalyzes the cleavage of para-nitroaniline (pNA) from a synthetic chromogenic substrate. The absorption of free pNA, determined at 405 nm, is inversely proportional to the activity of alpha-2-antiplasmin in plasma.
The process follows this scheme: Alpha-2-antiplasmin (plasma) + Plasmin (excess) => (Alpha-2-antiplasmin-plasmin) + Plasmin (residue). Substrate-pNA + Plasmin (residue) => Peptide + pNA.
A calibration curve is constructed using plasma calibrator with a known antiplasmin activity (A). Optical density values obtained for each dilution of the plasma calibrator are plotted on the Y-axis, and antiplasmin activity in % is plotted on the X-axis on a linear scale. Using the calibration curve and the optical density value of the test sample, the antiplasmin activity is determined.
Kit Composition:
• Plasmin, lyophilized – 2 vials;
• Plasma calibrator, lyophilized (1 ml/vial) – 1 vial;
• Chromogenic substrate, lyophilized (2 ml/vial) – 2 vials;
• Buffer solution (5 ml/vial) – 3 vials.
One kit is intended for conducting 20 macro determinations with a consumption of 0.5 ml of plasmin solution per analysis or for 100 micro determinations (tablet method) with a consumption of 0.075 ml of plasmin solution per analysis.
Results Interpretation: In the normal plasma of healthy individuals, the activity of antiplasmin is within the range of 80–120%.
Congenital deficiency of alpha-2-antiplasmin is associated with hemorrhagic conditions. A decrease in the level of alpha-2-antiplasmin is observed in liver diseases and disseminated intravascular coagulation (DIC) syndrome. An elevated level of alpha-2-antiplasmin is often observed in the postoperative period.
Normal and pathological values of alpha-2-antiplasmin should be monitored using control plasmas from RENAM Research and Production Company: Control Plasma, code KM-2.
