Method for detecting anticoagulant capacity of human prothrombin complex
A technology for human prothrombin and prothrombin is applied in the field of detecting the anticoagulation ability of human prothrombin complex, which can solve the problems of inconsistent content of anticoagulant components, difficulty in evaluating comprehensive anticoagulation ability, and inability to determine the risk of thrombosis. Easy operation and good repeatability
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Embodiment 1
[0031] The thrombin chromogenic substrate CS-01(38) was purchased from HYPEN-biomed Company, lot number: 30401-1. Embodiment 1 Detection method of the present invention
[0032] 1. Dilution of Human Prothrombin Complexes (PCCs) Preparations
[0033] Dilute the PCCs product with pure water to 0.8IU FIX / ml, and add 40μl to the microwell plate.
[0034] 2. PCCs mixed with human thrombin
[0035] Take 40 μl of human thrombin with a concentration of 4 IU / ml, add it to the above-mentioned microwell plate, shake slightly, mix well, and stand at room temperature for 1 min.
[0036] 3. Color reaction
[0037] Add 40 μl of the chromogenic substrate CS-01(38) (molecular formula: H-D-Phe-Pip-Arg-pNa·2HCl) solution at a concentration of 0.5 mg / ml to the above microwell plate, mix well, and incubate in a water bath at 22°C 4min.
[0038] 4. Absorbance measurement
[0039] The absorbance value (OD value) was measured at a wavelength of 405nm, once every 30s, and five times for a total ...
Embodiment 2
[0044] Embodiment 2 detection method of the present invention
[0045] 1. Dilution of Human Prothrombin Complexes (PCCs) Preparations
[0046] Dilute the PCCs product with pure water to 0.8-1.0 IU FIX / ml, and add 60 μl to the microwell plate.
[0047] 2. PCCs mixed with human thrombin
[0048] Take 60 μl of human thrombin with a concentration of 5 IU / ml, add it to the above-mentioned microwell plate, shake slightly, mix well, and place at room temperature for 1.5 min.
[0049] 3. Color reaction
[0050] Add 60 μl of the chromogenic substrate CS-01(38) (molecular formula: H-D-Phe-Pip-Arg-pNa·2HCl) solution at a concentration of 0.6 mg / ml to the above microwell plate, mix well, and incubate in a water bath at 25°C 5min.
[0051] 4. Absorbance measurement
[0052] The absorbance value (OD value) was measured at a wavelength of 405nm, once every 40s, and 7 times for a total of 280s.
[0053] 5. Establishment of linear equations
[0054] With the above-mentioned measured OD ...
Embodiment 3
[0057] Embodiment 3 detection method of the present invention
[0058] 1. Dilution of Human Prothrombin Complexes (PCCs) Preparations
[0059] Dilute the PCCs product with pure water to 1.3IU FIX / ml, and add 80μl to the microwell plate.
[0060] 2. PCCs mixed with human thrombin
[0061] Take 80 μl of human thrombin with a concentration of 6 IU / ml, add it to the above-mentioned microwell plate, shake slightly, mix well, and stand at room temperature for 2 minutes.
[0062] 3. Color reaction
[0063] Add 80 μl of the chromogenic substrate CS-01(38) (molecular formula: H-D-Phe-Pip-Arg-pNa 2HCl) solution with a concentration of 0.7 mg / ml to the above microwell plate, mix well, and incubate in a water bath at 28°C 6min.
[0064] 4. Absorbance measurement
[0065] The absorbance value (OD value) was measured at a wavelength of 405nm, once every 50 s, 8 times for a total of 400 s.
[0066] 5. Establishment of linear equations
[0067] With the above-mentioned measured OD valu...
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