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Kit for detecting content of neutrophil gelatinase-associated lipocalin (NGAL)

A technology of lipocalin and neutrophils, applied in the field of biological protein detection, can solve the problems of not being able to meet the wide linear range and high sensitivity at the same time, and inconvenient use

Active Publication Date: 2014-12-10
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 detecting content of neutrophil gelatinase-associated lipocalin (NGAL)
  • Kit for detecting content of neutrophil gelatinase-associated lipocalin (NGAL)
  • Kit for detecting content of neutrophil gelatinase-associated lipocalin (NGAL)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Reagent R1: pH7.4

[0074] Tris 50mM

[0075] NaCl 150mM

[0076] BSA 0.5%

[0077] Tween-20 0.1%

[0078] PEG6000 1.5%

[0079] NaN3 0.1%

[0080] Reagent R2: pH7.4

[0081] Tris 50mM

[0082] NaCl 150mM

[0083] BSA 0.5%

[0084] Tween-20 0.1%

[0085] NaN3 0.1%

[0086] Sucrose 5%

[0087] NGAL antibody sensitization latex particles 0.5%

[0088] NGAL reagent reference standard

[0089] Standard dilution buffer: pH7.4

[0090] Tris 50mM

[0091] NaCl 150mM

[0092] BSA 0.5%

[0093] Tween-20 0.1%

[0094] EDTA 2mM

[0095] NaN3 0.1%

[0096] Dissolve and dilute the recombinant human NGAL protein with the standard dilution solution according to the standard concentration (150, 300, 600, 1500, 3000, 5000ng / ml), and use the well-known domestic brand NGAL kit to measure and calibrate the prepared standard.

[0097]

[0098] The preparation process of NGAL antibody sensitized latex particles is as follows:

[0099] (1) Use 50mM MES buffer (pH 6.0) to...

Embodiment 2

[0106] Reagent R1: pH7

[0107] Tris 10mM

[0108] NaCl 125mM

[0109] BSA 0.05%

[0110] Tween-20 0.05%

[0111] PEG6000 1%

[0112] NaN3 0.1%

[0113] Reagent R2: pH7

[0114] Tris 20mM

[0115] NaCl 100mM

[0116] BSA 0.05%

[0117] Tween-20 0.08%

[0118] NaN 3 0.1%

[0119] Sucrose 2%

[0120] NGAL antibody sensitization latex particles 0.1%

[0121] NGAL reagent reference standard

[0122] Standard dilution buffer: pH7

[0123] Tris 40mM

[0124] NaCl 70mM

[0125] BSA 0.05%

[0126] Tween-20 0.03%

[0127] EDTA 5mM

[0128] NaN 3 0.1%

[0129] Dissolve and dilute the recombinant human NGAL protein with the standard dilution solution according to the standard concentration (150, 300, 600, 1500, 3000, 5000ng / ml), and use the well-known domestic brand NGAL kit to measure and calibrate the prepared standard.

[0130]

[0131] The preparation process of NGAL antibody sensitized latex particles is as follows:

[0132] (1) Use 50mM MES buffer (pH 6.0)...

Embodiment 3

[0139] Reagent R1: pH7.5

[0140] Tris 100mM

[0141]NaCl 50mM

[0142] BSA 1%

[0143] Tween-20 0.01%

[0144] PEG6000 0.5%

[0145] NaN3 0.1%

[0146] Reagent R2: pH7.5

[0147] Tris 100mM

[0148] NaCl 50mM

[0149] BSA 1%

[0150] Tween-20 0.01%

[0151] NaN 3 0.1%

[0152] Sucrose 10%

[0153] NGAL antibody sensitization latex particles 1%

[0154] NGAL reagent reference standard

[0155] Standard dilution buffer: pH7.5

[0156] Tris 10mM

[0157] NaCl 200mM

[0158] BSA 1%

[0159] Tween-20 0.01%

[0160] EDTA 0.5mM

[0161] NaN 3 0.1%

[0162] Dissolve and dilute the recombinant human NGAL protein with the standard dilution solution according to the standard concentration (150, 300, 600, 1500, 3000, 5000ng / ml), and use the well-known domestic brand NGAL kit to measure and calibrate the prepared standard.

[0163]

[0164] The preparation process of NGAL antibody sensitized latex particles is as follows:

[0165] (1) Use 50mM MES buffer (pH 6.0)...

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Abstract

The invention discloses a kit for detecting the content of neutrophil gelatinase-associated lipocalin (NGAL). The kit comprises a reagent R1, a reagent R2 and an NGAL reagent reference standard substance, wherein the reagent R1 comprises the following components: 10mM-100mM of Tris, 50mM-200mM of NaCl, 0.05%-1% of BSA (Bovine Serum Albumin), 0.01%-0.1% of Tween-20, 0.5%-3% of PEG (Polyethylene Glycol) and 0.1% of NaN3; the reagent R2 comprises the following components: 10mM-100mM of Tris, 50mM-200mM of NaCl, 0.05%-1% of BSA, 0.01%-0.1% of Tween-20, 0.1% of NaN3, 1%-10% of saccharose and 0.1%-1% of NGAL antibody sensitization latex particles; the NGAL reagent reference standard substance comprises the following components: 10mM-100mM of Tris, 50mM-200mM of NaCl, 0.05%-1% of BSA, 0.01%-0.1% of Tween-20, 0.5mM-5mM of EDTA (ethylenediamine tetraacetic acid) and 0.1% of NaN3. The kit is simple in reagent composition, high in test sensitivity, wide in linear range, high in stability, low in test cost, high in accuracy and convenient to popularize.

Description

[0001] technical field [0002] The invention relates to a biological protein detection technical solution, in particular to a neutrophil gelatinase-related lipocalin content detection kit. [0003] The meanings of the following expressions in this application are: [0004] 0.01-0.2M NaCl: indicates that the NaCl concentration in the solution is 0.01-0.2mol / L; [0005] 0.1-3% sucrose: means add 0.1-3g sucrose per 100mL solution. [0006] Tris: Trishydroxymethylaminomethane; [0007] BSA: bovine serum albumin; [0008] EDTA: ethylenediaminetetraacetic acid; [0009] EDAC: water-soluble carbodiimide crosslinking agent; [0010] PEG: polyethylene glycol; [0011] MES: Morpholinoethanesulfonic acid monohydrate [0012] The meanings expressed in this application that are similar to the above expressions are similar to the above meanings; unless otherwise stated in the application. [0013] Background technique [0014] Neutrophil gelatinase-associated lipocalin (NGAL...

Claims

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Application Information

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