Cup type time-resolved fluorescence BNP analysis method and reagent kit based on microspheres
A technology of time-resolved fluorescence and analysis method, which is applied in the field of cup-type time-resolved fluorescence BNP analysis method and kit based on microspheres, which can solve the problems of unsatisfactory detection precision, complicated detection steps, poor precision, etc., and reduce space Effects of steric hindrance, shorter detection time, and higher detection sensitivity
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Embodiment 1
[0058] 1. Preparation of detection cuvette
[0059] (1) Detection cuvette coating
[0060] The BNP antibody 50E1 (Hytest Company) was diluted to 20 μg / ml with 0.2M phosphate buffer (pH7.8), 200 μL / well, coated at 37° C. for 4 hours, and washed.
[0061] (2) Detect cuvette closure
[0062] Add 300 μL / well of blocking buffer to the reaction cup, block at 37°C for 4 hours, discard the blocking solution in the coated reaction cup, and pat dry.
[0063] (3) Detection of cuvette drying
[0064] The sealed cuvette was placed in a 37° C. drying oven with a humidity lower than 30% for 8 hours, sealed and dried for storage.
[0065] 2. Preparation of fluorescently labeled antibodies
[0066] (1) Pretreatment of fluorescent particles
[0067] Take 1mg of carboxyl time-resolved fluorescent microspheres (300nm, 0.1ml, 10mg / mL, Bangslab Company), add 60ul of 500mM MES (2-(N-morpholine)ethanesulfonic acid) buffer, pH 6.0, add 0.2mg of 1- (3-Dimethylaminopropyl)-3-ethylcarbodiimide hydr...
Embodiment 2
[0095] 1. Preparation of detection cuvette
[0096] (1) Detection cuvette coating
[0097] The BNP antibody 50E1 (Hytest Company) was diluted to 20 μg / ml with 0.2M phosphate buffer (pH7.8), 200 μL / well, coated at 37° C. for 4 hours, and washed.
[0098] (2) Detect cuvette closure
[0099] Add 300 μL / well of blocking buffer to the reaction cup, block at 37°C for 4 hours, discard the blocking solution in the coated reaction cup, and pat dry.
[0100] (3) Detection of cuvette drying
[0101] The sealed cuvette was placed in a 37° C. drying oven with a humidity lower than 30% for 8 hours, sealed and dried for storage.
[0102] 2. Preparation of fluorescently labeled antibodies
[0103] (1) Pretreatment of fluorescent particles
[0104] Take 1mg of carboxyl time-resolved fluorescent microspheres (300nm, 0.1ml, 10mg / mL, Bangslab Company), add 60ul of 500mM MES (2-(N-morpholine)ethanesulfonic acid) buffer, pH 6.0, add 0.2mg of 1- (3-Dimethylaminopropyl)-3-ethylcarbodiimide hydr...
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