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Enzyme-linked immunosorbent assay chromogenic substrate solution

A chromogenic substrate and enzyme-linked immunosorbent technology, which is applied in biological testing, material inspection products, and analysis by making materials undergo chemical reactions, can solve problems such as difficult storage, reduced color intensity, and unstable properties, and achieve Enhanced color intensity, improved sensitivity, and enhanced uniformity

Active Publication Date: 2014-03-19
BIOBASE BIODUSTRY (SHANDONG) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, TMB is relatively difficult to dissolve in water, and its properties are unstable in the solution state, so it is difficult to store, which results in the poor stability of the conventional TMB chromogenic solution on the market, the color development is not durable enough, and the color intensity decreases after a long period of storage. question

Method used

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  • Enzyme-linked immunosorbent assay chromogenic substrate solution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: Preparation of TMB chromogenic substrate solution

[0030] Chromogenic solution A:

[0031] Sodium citrate 20g / L

[0032] Citric acid 10 g / L

[0033] Carbamide peroxide 1g / L

[0034] Sodium pyrophosphate 5g / L

[0035] Hydrogen peroxide 1mL / L

[0036] Polyoxyethylene lauryl ether 01mL / L

[0037] N-Methylisothiazolone-HCl (MIT) 10g / L

[0038] Each component was dissolved in 1L of distilled water to complete the preparation;

[0039] Chromogenic solution B:

[0040] TMB 5g / L

[0041] Citric acid 10 g / L

[0042] Disodium EDTA 2 g / L

[0043] Glycerin 10 mL / L

[0044] Dimethyl sulfoxide 10 mL / L

[0045] Dissolve TMB in dimethyl sulfoxide first, then add 1L of distilled water, and other components are sequentially added and dissolved.

[0046]The chromogenic substrate solution was tested and verified by the hepatitis C core antigen detection kit (double-antibody sandwich ELISA). The specific operation steps are: (1) Set up the sample well, add 50 μL...

Embodiment 2

[0051] Embodiment 2: prepare TMB chromogenic substrate solution as follows,

[0052] Chromogenic solution A:

[0053] Sodium citrate 5g / L

[0054] Citric acid 2 g / L

[0055] Carbamide peroxide 0.5g / L

[0056] Sodium pyrophosphate 0.5g / L

[0057] Hydrogen peroxide 0.5mL / L

[0058] Polyoxyethylene lauryl ether 0.2mL / L

[0059] N-Methylisothiazolone-HCl (MIT) 1g / L

[0060] Each component was dissolved in 1L of distilled water to complete the preparation.

[0061] Chromogenic solution B:

[0062] TMB 0.3g / L

[0063] Citric acid 2 g / L

[0064] Disodium edetate 0.2g / L

[0065] Glycerin 1mL / L

[0066] Dimethyl sulfoxide 5 mL / L

[0067] Dissolve TMB in dimethyl sulfoxide first, then add 1L of distilled water, and other components are sequentially added and dissolved.

[0068] The chromogenic substrate solution prepared in Example 2 and the common chromogenic substrate solution were placed in an incubator at 37°C at the same time. After 7 days, they were taken out at the s...

Embodiment 3

[0072] Embodiment 3: prepare TMB chromogenic substrate solution as follows,

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Abstract

The invention relates to an enzyme-linked immunosorbent assay substrate solution. The substrate solution comprises a chromogenic solution (A) and a chromogenic solution (B). The substrate solution provided by the invention can be stably stored for two years at the temperature of 2 to 8 DEG C, the chromogenic effect can be kept for 1h, and the chromogenic strength is obviously improved; meanwhile, a background is not added, so that the detection sensitivity is improved, the aims of stability and high efficiency are achieved, and various requirements of an enzyme-linked immunosorbent assay kit on the chromogenic substrate solution are met.

Description

technical field [0001] The invention relates to a stable and efficient enzyme-linked immunosorbent substrate chromogenic solution, which belongs to the technical field of clinical in vitro detection. Background technique [0002] In 1971 Engvall and Perlmann published ELISA Adsorbent Determination (enzyme linked immunosorbent assay, Elis A) The article used for the quantitative determination of IgG made the enzyme-labeled antibody technology used for antigen localization in 1966 develop into a method for the determination of trace substances in liquid specimens. The basics of this method principle It is: ① Make the antigen or antibody bind to the surface of a certain solid phase carrier and maintain its immunological activity. ②The antigen or antibody is linked with horseradish peroxidase (HRP) to form an enzyme-labeled antigen or antibody, which not only retains its immune activity, but also retains the activity of the enzyme. During the determination, the test specim...

Claims

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Application Information

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IPC IPC(8): G01N21/78G01N33/52
Inventor 谭柏清王进甘宜梧
Owner BIOBASE BIODUSTRY (SHANDONG) CO LTD
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