Method for measuring activity of enzyme for dissolving thrombus
A determination method and thrombolytic enzyme technology are applied in the direction of determination/inspection of microorganisms, biochemical equipment and methods, etc., which can solve the problems of cumbersome thrombus preparation, low determination efficiency, outdated and backward, etc., to shorten the determination time and improve the determination efficiency. , high precision and accuracy
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experiment Embodiment 1
[0021] Experimental Example 1: Urokinase Standard Curve Determination 1
[0022] To make microthrombosis: use 0.04mol / L, pH7.8 sodium barbital buffer solution to prepare fibrinogen solution with a concentration of 6.7mg / mL, a thrombin solution with a concentration of 300u / mL, and a plasma solution with a concentration of 1 casein unit / mL. Enzyme solution, 1.28% (w / v) agarose solution, add thrombin solution to the agarose solution so that the final concentration of thrombin is 3.6u / mL, the specific operations are as follows:
[0023] Take a 50ml test tube, add 2.5ml of agarose solution to the test tube, place the test tube in a water bath at 40-42°C, add 35 μL of thrombin solution to the test tube, add 50 μL of plasma lysinogen solution of 1 casein unit / mL , shake and mix well, then add 1.5mL of fibrinogen solution, mix quickly and add 150μL to the small well of the cell culture plate, incubate the plate in a humid box at 37°C for 1 hour to make a microthrombosis plate; (If th...
experiment Embodiment 2
[0034] Experimental Example 2: Urokinase Standard Curve Determination 2
[0035] When making microthrombosis, the concentration of the agarose solution was prepared to be 1.6%, and the rest of the steps were the same as in Experimental Example 1. can be made image 3 Urokinase standard curve, the standard curve fitting equation is y=0.022x+1.3656, r 2 =0.9904
experiment Embodiment 3
[0036] Experimental Example 3: Urokinase Standard Curve Determination 3
[0037] When making microthrombosis, the concentration of the agarose solution was prepared to be 0.96%, and the rest of the steps were the same as in Experimental Example 1. can be made Figure 4 Urokinase standard curve, the standard curve fitting equation is y=0.0208x+1.3648, r 2 =0.9801
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