Neutrophil gelatinase-associated lipocalin determination kit
A technology of lipocalin and neutrophils, which is applied in the fields of medicine and biochemistry, can solve the problems of low measurement accuracy, radioactive pollution, poor stability, etc., achieve sensitive and accurate detection results, and remove the interference of rheumatic factors Effect
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Embodiment 1
[0082] Reagent R1:
[0083] Tris buffer 50 mmol / L
[0084] Potassium chloride 100mmol / L
[0085] Polyethylene glycol-8000 20g / L
[0086] Sodium Potassium Tartrate 45mmol / L
[0087] Bovine serum albumin 20 g / L
[0088] Sodium Benzoate 0.8g / L
[0089] Goat anti-human polyclonal antibody 0.8 g / L
[0090] The solvent is purified water
[0091] Reagent R2:
[0092] Tris buffer 50 mmol / L
[0093] Bovine Serum Albumin 17 g / L
[0094] Glycerin 20 mmol / L
[0095] Triton-100 1.0mL / L
[0096] Phenol 0.7g / L
[0097] Latex coated anti-human neutrophil gelatinase-associated lipocalin 1.0 g / L
[0098] Its solvent is purified water.
Embodiment 2
[0100] The kit of the present invention includes reagent R1 and reagent R2 two-liquid components independent of each other, wherein
[0101] Reagent R1:
[0102] MOPS buffer 80 mmol / L
[0103] Magnesium chloride 50mmol / L
[0104] Polyethylene glycol-2000 30 g / L
[0105] Sodium Potassium Tartrate 25 mmol / L
[0106] Trehalose 30 g / L
[0107] Sodium azide 0.4 g / L
[0108] Goat anti-human monoclonal antibody 1.4 g / L
[0109] The solvent is purified water
[0110] Reagent R2:
[0111] MOPS buffer 80 mmol / L
[0112] Trehalose 12 g / L
[0113] Sodium alginate 35 mmol / L
[0114] Triton-100 0.5 mL / L
[0115] Sodium azide 0.9 g / L
[0116] Latex coated anti-human neutrophil gelatinase-associated lipocalin 0.2 g / L
[0117] Its solvent is purified water.
Embodiment 3
[0119] Kit preparation and method of use
[0120] 1. Preparation of latex-coated anti-human neutrophil gelatinase-associated lipocalin: use 50 mmol / L MES buffer to dilute polystyrene microspheres with a particle size of 120 nm into polystyrene microspheres contained in them. The mass concentration of spheres is 3% solution, then add 0.5 mg 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride to each milliliter solution, and react at 28°C For 2 hours, use a 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 at 15,000 rpm / min centrifuge for 30 minutes at a high speed, remove the supernatant, suspend the pellet in 50 mmol / L MES buffer, disperse by ultrasonic, and then add an equal volume of anti-human neutrophil gelatinase-associated lipocalin while stirring MES dilution of antibody, mixed and stirred, reacted at 26°C for 2 hours, so that the final concentrati...
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