Kit for content detection of neutrophil gelatinase-associated lipocalin

A technology for lipocalin and neutrophils, applied in the field of biological protein detection, can solve the problems of inability to meet wide linear range and high sensitivity at the same time, inconvenient to use, etc.

Active Publication Date: 2015-12-23
NINGBO RUI BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are kits for the determination of NGAL concentration in human plasma or urine by latex-enhanced immunoturbidimetry, but these kits cannot meet the requirements of wide linear range and high sensitivity at the same time, so it is necessary to dilute the sample to measure high concentration of NGAL , which is inconvenient to use

Method used

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  • Kit for content detection of neutrophil gelatinase-associated lipocalin
  • Kit for content detection of neutrophil gelatinase-associated lipocalin
  • Kit for content detection of neutrophil gelatinase-associated lipocalin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] In the present embodiment, in the carbonate buffer solution of PH8, add SDS, KPS and VN monomer, the content of VN monomer in carbonate buffer solution is 3wt%, the content of KPS in carbonate buffer solution is 0.1wt%, the content of SDS in the carbonate buffer solution is 0.15wt%, after stirring the reaction under normal pressure protective gas atmosphere and water bath conditions, the temperature of the water bath is 83 degrees Celsius, and the protective gas atmosphere is a mixture of nitrogen and carbon dioxide, Among them, carbon dioxide accounts for 15 (v / v)% of the total amount of the mixed gas. After the reaction is completed, after high-speed separation by the centrifuge, wash with deionized water or double distilled water for 1-2 times, then wash with deionized water or double distilled water and separate with a semi-permeable membrane to obtain PVN nano latex particles. The particle size of PVN nano latex particles is 80nm.

Embodiment 2

[0064] In this embodiment, in the carbonate buffer solution of pH8.5, add SDS, KPS and VN monomer, the content of VN monomer in the carbonate buffer solution is 4wt%, the content of KPS in the carbonate buffer solution The content is 0.2wt%, and the content of SDS in the carbonate buffer is 0.08wt%. After stirring and reacting under normal pressure protective gas atmosphere and water bath conditions, the temperature of the water bath is 78 degrees Celsius, and the protective gas atmosphere is a mixture of nitrogen and carbon dioxide. Gas, in which carbon dioxide accounts for 12 (v / v)% of the total mixture. After the reaction is completed, after high-speed separation by the centrifuge, wash with deionized water or double distilled water for 1-2 times, then wash with deionized water or double distilled water and separate with a semi-permeable membrane to obtain PVN nano latex particles. The particle size of PVN nano latex particles is 120nm.

Embodiment 3

[0066] In the present embodiment, in the carbonate buffer solution of PH9, add SDS, KPS and VN monomer, the content of VN monomer in carbonate buffer solution is 5wt%, the content of KPS in carbonate buffer solution is 0.3wt%, the content of SDS in the carbonate buffer solution is 0.10wt%, after stirring the reaction under normal pressure protective gas atmosphere and water bath conditions, the temperature of the water bath is 75 degrees Celsius, and the protective gas atmosphere is a mixture of nitrogen and carbon dioxide, Among them, carbon dioxide accounts for 10 (v / v)% of the total amount of the mixed gas. After the reaction is completed, after high-speed separation by the centrifuge, wash with deionized water or double distilled water for 1-2 times, then wash with deionized water or double distilled water and separate with a semi-permeable membrane to obtain PVN nano latex particles. The particle size of PVN nano latex particles is 110nm.

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Abstract

The invention discloses a kit for content detection of neutrophil gelatinase-associated lipocalin, wherein a reagent R1 comprises Tris, NaCl, BSA, Tween-20, PEG and NaN3; a reagent R2 comprises Tris, NaCl, BSA, Tween-20, NaN3, sucrose and NGAL antibody sensitized latex particles; a NGAL reagent reference standard sample comprises Tris, NaCl, BSA, Tween-20, EDTA, NaN3 and NGAL of different contents; the NGAL antibody sensitized latex particles comprise PS nanometer latex particles and PVN nanometer latex particles, and the particle size of the PS nanometer latex particle is larger than the particle size of the PVN nanometer latex particle. The invention has the advantages of simple reagent composition, good test sensitivity, wide linearity range, good stability, low test cost and high precision, and the kit is convenient for popularization.

Description

technical field [0001] The invention relates to a biological protein detection technical solution, in particular to a neutrophil gelatinase-related lipocalin content detection kit. [0002] The meanings of the following expressions in this application are: [0003] 0.01-0.2MNaCl: indicates that the NaCl concentration in the solution is 0.01-0.2mol / L; [0004] 0.1-3% sucrose: means adding 0.1-3g sucrose per 100mL solution. [0005] Tris: Trishydroxymethylaminomethane; [0006] BSA: bovine serum albumin; [0007] EDTA: ethylenediaminetetraacetic acid; [0008] EDAC: water-soluble carbodiimide crosslinking agent; [0009] PEG: polyethylene glycol; [0010] MES: Morpholineethanesulfonic acid monohydrate; [0011] PVN refers to polyvinyl naphthalene, VN refers to its monomer; SDS refers to sodium dodecyl sulfonate; KPS refers to potassium persulfate; [0012] The meanings expressed in this application that are similar to the above expressions are similar to the above meanin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 陈媛许琴张闻周海滨王建飞
Owner NINGBO RUI BIO TECH
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