A kind of cystine protease inhibitor c detection kit and its detection method
A cysteine protease and detection kit technology, which is applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems of poor repeatability and cumbersome operation, and achieve the effects of low cost, high detection sensitivity and good adaptability
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Embodiment 1
[0035] Example 1 Preparation of Latex Particle Anti-Human Cys C Antibody Conjugated Emulsion
[0036] The preparation method of latex particle anti-human Cys C antibody binding emulsion comprises the steps:
[0037] 1) Take 1 mL of carboxylated latex microspheres with a particle size of 120 nm and a solid content of 10%, add glycine buffer to 5 mL;
[0038] 2) Add 1mL of 22g / L N-hydroxysuccinimide and 1mL of 8.5g / L 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride for activation, stir After uniformity, the first incubation was carried out at a temperature of 37°C for 20 minutes to obtain activated polystyrene latex microsphere particles;
[0039] 3) Add 3 mL of phosphate buffer, stir at room temperature at a speed of 300 r / min, and add anti-human Cys C at a mass ratio of anti-human Cys C monoclonal antibody to the polystyrene latex microsphere particles of 0.04:1. Monoclonal antibody, for the second incubation, the incubation temperature is 37°C, and the incubation...
Embodiment 2
[0041] Example 2 Preparation of Latex Particle Anti-Human Cys C Antibody Conjugated Emulsion
[0042] The preparation method of latex particle anti-human Cys C antibody binding emulsion comprises the steps:
[0043] 1) Take 2mL of carboxylated latex microspheres with a particle size of 120nm and a solid content of 10%, add glycine buffer to 5mL;
[0044] 2) Add 3mL of 22g / L N-hydroxysuccinimide and 3mL of 8.5g / L 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride for activation, stir After uniformity, the first incubation was carried out at a temperature of 37°C for 20 minutes to obtain activated polystyrene latex microsphere particles;
[0045] 3) Add 6 mL of phosphate buffer, stir at room temperature at a speed of 300 r / min, and add anti-human Cys C at a mass ratio of anti-human Cys C monoclonal antibody to the polystyrene latex microsphere particles at a ratio of 0.1:1. Monoclonal antibody, for the second incubation, the incubation temperature is 37°C, and the inc...
Embodiment 3
[0047] Example 3 Cystatin C detection kit and detection method
[0048] The cystatin C detection kit includes reagent R1 and reagent R2; the volume ratio of reagent R1 to reagent R2 is 4:1.
[0049] R1: Glycine buffer 50mmoL / L, NaCl 25g / L, polyethylene glycol 8000 8g / L, Tween-20 30g / L, Proclin-300 0.5g / L;
[0050] R2: Phosphate buffer 150mmoL / L, latex particle anti-human Cys C antibody-conjugated emulsion 0.1g / L prepared in Example 1, Tween-20 10g / L, bovine serum albumin (BSA) 20g / L, glycerin 80g / L, Betaine 10g / L, Proclin3000.5g / L.
[0051] The detection method of the cystatin C detection kit includes the following steps: separate the serum sample, add reagent R1 to the serum sample, mix well, incubate for 4 minutes, add R2, mix well and incubate for 25 seconds, then detect at a wavelength of 600nm Absorbance value A1, after 5 minutes, detect absorbance value A2 at a wavelength of 700nm, establish a standard curve based on serum standard data, and calculate the content of c...
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