Cup type time-resolved fluorescence FABP analysis method and reagent kit based on microspheres
A time-resolved fluorescence and analysis method technology, applied in the field of cup-type time-resolved fluorescence FABP analysis methods and kits based on microspheres, can solve the problems of unsatisfactory precision, complicated detection steps, poor precision, etc. The effect of reducing the resistance effect, shortening the detection time, and improving the detection sensitivity
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Embodiment 1
[0057] 1. Preparation of detection cuvette
[0058] (1) Detection cuvette coating
[0059] The h-FABP antibody 2302 (Medix Company) was diluted to 20 μg / ml with 0.2M phosphate buffer (pH 7.8), 200 μL / well, coated at 37° C. for 4 hours, and the plate was washed.
[0060] (2) Detect cuvette closure
[0061] 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.
[0062] (3) Detection of cuvette drying
[0063] 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.
[0064] 2. Preparation of fluorescently labeled antibodies
[0065] (1) Pretreatment of fluorescent particles
[0066] Take 1mg of carboxyl time-resolved fluorescent microspheres (300nm, 0.1ml, 10mg / mL, Bangslab company), add 60ul 500mM MES (2-(N-morpholine)ethanesulfonic acid) buffer, pH 6.0, add 0.2mg 1 -(3-Dimethylaminopropyl)-3-ethylcarbod...
Embodiment 2
[0093] 1. Preparation of detection cuvette
[0094] (1) Detection cuvette coating
[0095] The h-FABP antibody 2302 (Medix Company) was diluted to 20 μg / ml with 0.2M phosphate buffer (pH 7.8), 200 μL / well, coated at 37° C. for 4 hours, and the plate was washed.
[0096] (2) Detect cuvette closure
[0097] 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.
[0098] (3) Detection of cuvette drying
[0099] 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.
[0100] 2. Preparation of fluorescently labeled antibodies
[0101] (1) Pretreatment of fluorescent particles
[0102] Take 1mg of carboxyl time-resolved fluorescent microspheres (300nm, 0.1ml, 10mg / mL, Bangslab Company), add 60ul 500mM MES (2-(N-morpholine)ethanesulfonic acid) buffer, pH 6.0, add 0.2mg 1 -(3-Dimethylaminopropyl)-3-ethylcarbod...
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