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Preparation and formula of single-component horse radish peroxidase (HRP) substrate (TMB) color developing solution

A chromogenic liquid and one-component technology, which is applied in the direction of material analysis, measuring devices, instruments, etc. by observing the influence of chemical indicators, can solve problems such as poor uniformity, poor stability, and inconvenient operation, and achieve Improves water solubility, prevents oxidation, and has the effect of long signal stabilization time

Inactive Publication Date: 2019-05-17
伊艾博(武汉)科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The TMB chromogen solution used in the existing commercially available enzyme-linked immunosorbent kits is divided into two types, one is two-component, divided into A solution and B solution, and it needs to be mixed before use, because it is easy to cause manipulation during use. Inconvenient and short storage time after mixing are gradually replaced by the second type; the second type is one-component TMB, which is easy to operate, but the single-component TMB currently used in the market has poor stability and short storage time. Most of the storage time is 3-6 months, and the color background is high, and the uniformity is not good

Method used

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  • Preparation and formula of single-component horse radish peroxidase (HRP) substrate (TMB) color developing solution
  • Preparation and formula of single-component horse radish peroxidase (HRP) substrate (TMB) color developing solution
  • Preparation and formula of single-component horse radish peroxidase (HRP) substrate (TMB) color developing solution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] An embodiment of the one-component TMB chromogenic solution of the present invention, the one-component TMB chromogenic solution described in this embodiment comprises the following components: TMB, Tween 80, hydroxypropyl-β-cyclodextrin, L-formazol Thionine, DMSO, disodium hydrogen phosphate, citric acid, hydrogen peroxide; Wherein, the mass concentration of the TMB in the one-component TMB color developing solution is 0.1g / L, and the Tween 80 is in the The mass concentration in the one-component TMB chromogenic solution is 4g / L, and the mass concentration of the hydroxypropyl-β-cyclodextrin in the one-component TMB chromogenic solution is 2.5g / L, and the The mass concentration of L-methionine in the one-component TMB chromogenic solution is 0.3g / L, and the volume concentration of the DMSO in the one-component TMB chromogenic solution is 20ml / L, and the The molar concentration of disodium hydrogen phosphate in the one-component TMB chromogenic solution is 0.1mol / L, and...

Embodiment 2

[0052] An embodiment of the one-component TMB chromogenic solution of the present invention, the one-component TMB chromogenic solution described in this embodiment comprises the following components: TMB, Tween 80, hydroxypropyl-β-cyclodextrin, L-formazol Thionine, DMSO, disodium hydrogen phosphate, citric acid, hydrogen peroxide; Wherein, the mass concentration of described TMB in described one-component TMB chromogenic solution is 0.2g / L, and described Tween 80 is in the The mass concentration in the one-component TMB chromogenic solution is 6g / L, and the mass concentration of the hydroxypropyl-β-cyclodextrin in the one-component TMB chromogenic solution is 4g / L, and the L - the mass concentration of methionine in the one-component TMB chromogenic solution is 0.2g / L, the volume concentration of the DMSO in the one-component TMB chromogenic solution is 15ml / L, and the phosphoric acid The molar concentration of disodium hydrogen in the one-component TMB chromogenic solution i...

Embodiment 3

[0055] An embodiment of the one-component TMB chromogenic solution of the present invention, the one-component TMB chromogenic solution described in this embodiment comprises the following components: TMB, Tween 80, hydroxypropyl-β-cyclodextrin, L-formazol Thionine, DMSO, disodium hydrogen phosphate, citric acid, hydrogen peroxide; Wherein, the mass concentration of the TMB in the single-component TMB chromogenic solution is 0.4g / L, and the Tween 80 in the The mass concentration in the one-component TMB chromogenic solution is 10g / L, and the mass concentration of the hydroxypropyl-β-cyclodextrin in the one-component TMB chromogenic solution is 5g / L, and the L - the mass concentration of methionine in the one-component TMB chromogenic solution is 0.1g / L, the volume concentration of the DMSO in the one-component TMB chromogenic solution is 10ml / L, and the phosphoric acid The molar concentration of disodium hydrogen in the one-component TMB chromogenic solution is 0.05mol / L, and ...

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Abstract

The invention discloses a horse radish peroxidase (HRP) substrate color developing solution (single-component TMB solution). The component further comprises some kinds of organic antioxidant ascorbicacid (VC), antioxidative peptide and the like. The situation that color developing performance is affected by an unstable system caused by oxidation of the TMB solution in the storage and transportation process can be prevented. The single-component TMB color developing solution can be stably stored for more than twelve months, and meanwhile, the color developing performance of the single-component TMB color developing solution is better compared with a market double-component TMB color developing solution.

Description

technical field [0001] The invention relates to a biological in vitro diagnostic reagent and a preparation method thereof, in particular to a TMB chromogenic solution and a preparation method thereof. Background technique [0002] TMB (3,3',5,5'-tetramethylbenzidine) is a new type of safe chromogen reagent, as a new substrate of peroxidase, obtained in enzyme-linked immunosorbent assay (ELISA) widely used. The substrate yields a soluble light blue end product that can be read on a spectrophotometer at 370 or 630 nm. It is suitable for quantitative and qualitative analysis in the chromogenic stage of ELISA. The principle of the test is: when the enzyme labeled on the antibody or antigen reacts with TMB, hydrogen peroxide, etc. in the chromogenic solution, the solution turns from colorless to yellow after adding the stop solution, and the depth of yellow reflects the amount of antigen-antibody binding in the ELISA system , so as to judge the amount of a certain antigen or a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N33/531
Inventor 徐世瑞李学斌
Owner 伊艾博(武汉)科技股份有限公司
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