Retinol binding protein determination kit and preparation method thereof
A protein and retinol-binding technology, applied in the fields of medicine and biochemistry, can solve the problems of complex operation, radioactive contamination, and poor accuracy, and achieve the effects of increased solubility, high accuracy, and rapid response
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Embodiment 1
[0073] The kit of the present invention includes reagent R1 and reagent R2 two-liquid components independent of each other, wherein
[0074] Reagent R1:
[0075] Tris buffer 100 mmol / L
[0076] Dithizone 30 mmol / L
[0077] Polyethylene glycol-8000 20 g / L
[0078] Bovine Serum Albumin 40 g / L
[0080] The solvent is purified water
[0081] Reagent R2:
[0082] Tris buffer 100 mmol / L
[0083] Bovine serum albumin 12g / L
[0084] Triton-100 0.7 mL / L
[0085] Glycerin 45 mmol / L
[0087] Latex coated anti-human retinol binding protein antibody 4g / L
[0088] Its solvent is purified water.
Embodiment 2
[0090] The kit of the present invention includes reagent R1 and reagent R2 two-liquid components independent of each other, wherein
[0091] Reagent R1:
[0092] PBS buffer 150 mmol / L
[0093] 8-hydroxyquinoline 10 mmol / L
[0094] Polyethylene glycol-6000 30 g / L
[0095] Sucrose 20g / L
[0096] Phenol 1.1 g / L
[0097] The solvent is purified water
[0098] Reagent R2:
[0099] PBS buffer 50mmol / L
[0100] Bovine serum albumin 18 g / L
[0101] Triton-100 1.0 mL / L
[0102] Sodium alginate 60 mmol / L
[0103] Phenol 0.3 g / L
[0104] Latex coated anti-human retinol binding protein antibody 8g / L
[0105] Its solvent is purified water.
Embodiment 3
[0107] Kit preparation and method of use
[0108] 1. Preparation of latex-coated anti-human retinol-binding protein antibody: Dilute polystyrene microspheres with a particle size of 120 nm with 50 mmol / L MES buffer solution to obtain a mass concentration of polystyrene microspheres of 3% solution, and then add 0.5 mg of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride to each milliliter of the solution, react at 25°C for 2 hours, use Centrifuge at 15,000 rpm / min for 30 minutes, remove the supernatant, suspend the pellet in 50 mmol / L MES buffer, ultrasonically disperse, and then use a centrifuge to centrifuge at 15,000 rpm / min for 30 minutes Minutes, remove the supernatant, suspend the precipitate in 50 mmol / L MES buffer, make the final mass concentration of polystyrene microspheres 2%, ultrasonically disperse, and then add an equal volume of anti-human retinol containing The MES buffer solution bound to the protein antibody was mixed and stirred, and reacted at 25...
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