Neutrophil gelatinase-associated lipocalin (NGAL) protein level ELISA kit

A technology of lipocalin and neutrophils, which is applied in the field of biological protein detection, can solve the problems of reducing false negatives and increasing false positive results, and achieves the effect of increasing the dissociation speed

Active Publication Date: 2017-06-09
NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method is likely to increase the false positive results of non-specific reactions to free MMP-9; in additi

Method used

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  • Neutrophil gelatinase-associated lipocalin (NGAL) protein level ELISA kit
  • Neutrophil gelatinase-associated lipocalin (NGAL) protein level ELISA kit
  • Neutrophil gelatinase-associated lipocalin (NGAL) protein level ELISA kit

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preparation example Construction

[0037] 1. Preparation of carboxylated activated polystyrene latex solution: take 0.1g of polystyrene latex particles ((the polystyrene latex particles are a commercially available conventional product)) with a particle diameter of 150nm and add 10mL of 0.1M phosphoric acid Salt buffer (pH7.5) was dispersed, then 4 mg of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) was added, and 2 mg of NHS was added, and the reaction was stirred at room temperature for 0.5 hours , centrifuge and discard the supernatant to retain the precipitate, then dilute the dispersed precipitated latex with 10mL of 0.1M phosphate buffer to obtain the carboxylated modified polystyrene latex;

[0038] 2. Preparation of NGAL antibody solution: Dissolve 250 μg NGAL antibody in 0.5 mL deionized water to obtain NGAL antibody solution;

[0039] 3. Preparation of latex particles coated with NGAL antibody: Take 100 μL of the NGAL antibody solution prepared in the second step, dilute it with 5 mL of 0.1M ph...

Embodiment 1

[0041] 1. The preparation method of reagent Ⅰ:

[0042] Prepare reagent Ⅰ according to the following table:

[0043] Reagent Ⅰ concentration Tris(Tris) 0.1mol / L Guanidine Hydrochloride 0.1mmol / L

[0044] Use NaOH or HCl to adjust the pH value of Agent I to 7.4.

[0045] 2. Preparation method of reagent II:

[0046] Prepare reagent II according to the following table:

[0047]

[0048] Use NaOH or HCl to adjust the pH value of II to 7.0.

Embodiment 2

[0050] 1. The preparation method of reagent Ⅰ:

[0051] Prepare reagent Ⅰ according to the following table:

[0052]

[0053] Adjust the pH to 7.4 with NaOH or HCl.

[0054] 2. Preparation method of reagent II:

[0055] Prepare reagent II according to the following table:

[0056]

[0057]

[0058] Adjust the pH to 7.0 with NaOH or HCl.

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Abstract

Disclosed is a neutrophil gelatinase-associated lipocalin (NGAL) protein level ELISA kit which is composed of a reagent I and a reagent II. The reagent I comprises a slow-release agent and a denaturant; the reagent II comprises latex particles coated with NGAL antibodies. Aggregated protein is denatured to some extent after being added with the denaturant, physical and (or) chemical binding site is exposed, chemical binding action of the chemical binding site is fractured through sulfydryl dissociation agent , while physical binding action of the physical binding site is dispersed by surface active agent, so that the aggregated protein is disaggregated. Therefore, it is quite important to choose the appropriate types and concentrations of denaturant, sulfydryl dissociation agent, and surface active agent; the aggregate is disaggregated without interference on following immunological detection. According to the method, the the appropriate types and concentrations of denaturant, sulfydryl dissociation agent and surface active agent are determined and chosen specifically, which solves the technical problem above.

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. Background technique [0002] Human neutrophil gelatinase-associated lipocalin (neutrophilgelatinase-associated lipocalin, NGAL) molecular weight is about 25kDa, covalently bound to neutrophil gelatinase. NGAL is a growth factor in physiological state, mainly involved in the occurrence and growth of early kidney epithelium. The expression level is low in normal tissues including kidney, lung, stomach and colon epithelial tissues. After renal ischemia-reperfusion injury, the epithelium of the proximal convoluted tubule expresses a large amount of NGAL, which is a recently discovered sensitive biomarker for the early diagnosis of AKI (acute kidney injury). Urinary NGAL is a sensitive indicator of renal ischemia-reperfusion injury, and its expression level is correlated with renal ischemia time. ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N21/31
CPCG01N21/31G01N33/6893G01N2021/3129
Inventor 邹炳德邹继华方亮
Owner NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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