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

Inactive Publication Date: 2016-11-09
ANHUI IPROCOM BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The technical problem to be solved by the present invention is to provide a kit for measuring neutrophil gelatinase-associat

Method used

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  • Neutrophil gelatinase-associated lipocalin determination kit

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a neutrophil gelatinase-associated lipocalin determination kit. The kit comprises a reagent R1 and a reagent R2 which are liquid components and are mutually independent, the reagent R1 comprises a buffer solution, inorganic salt ions, an accelerator, a chelating agent, a stabilizer, an antiseptic and an anti-humanr heumatoid factor antibody, and the reagent R2 comprises the buffer solution, the stabilizer, a suspending aid, a surfactant, the antiseptic and a latex coated anti-human neutrophil gelatinase-associated lipocalin. The preparation method comprises the following steps: preparing the reagents according to the component content; mixing a sample to be determined with the reagent R1 and the reagent R2 to fully react the sample to be determined with the reagent R1 and the reagent R2; determining the absorbance difference by using a fully-automatic biochemical analyzer after the reaction; and calculating the concentration of neutrophil gelatinase-associated lipocalin in the sample according to the absorbance change value. The kit has high accuracy.

Description

technical field [0001] The invention relates to the technical fields of medicine and biochemistry, in particular to a kit for measuring neutrophil gelatinase-associated lipocalin. Background technique [0002] Neutrophil gelatinase-associated lipocalin (NGAL) is a kind of lipocalin. It was originally a small molecular weight secreted protein found in activated neutrophils. Modern studies have shown that NGAL is a diagnostic tool for acute kidney injury. One of the most effective biological markers, and also one of the effective markers of early diabetic nephropathy. [0003] Neutrophil gelatinase-associated lipocalin (NGAL) has powerful functions. In addition to binding and transporting small hydrophobic molecules as a member of the lipid family, it is also associated with inflammation, embryonic development, immune response, and chemotaxis , signal transduction, and the occurrence and development of various tumors. [0004] Neutrophil gelatinase-associated lipocalin (NGAL...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6893G01N2800/042G01N2800/347
Inventor 蔡晓辉庄庆华吴铮徐运
Owner ANHUI IPROCOM BIOTECH CO LTD
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