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Enhanced chemiluminiscence kit and preparation method of enhanced chemiluminiscence kit

A chemiluminescent reagent and enhanced technology, which is applied in chemiluminescence/bioluminescence, analysis through chemical reaction of materials, biological testing, etc., can solve the problems of poor stability of hydrogen peroxide, short product storage time, serious degradation, etc. problems, to achieve high stability, high thermal stability, strong oxygen supply effect

Inactive Publication Date: 2018-01-09
JIANGSU KEYGEN BIOTECH CORP LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, hydrogen peroxide has poor stability, short storage time as a product, severe degradation during transportation, and unstable quality

Method used

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  • Enhanced chemiluminiscence kit and preparation method of enhanced chemiluminiscence kit
  • Enhanced chemiluminiscence kit and preparation method of enhanced chemiluminiscence kit
  • Enhanced chemiluminiscence kit and preparation method of enhanced chemiluminiscence kit

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Embodiment Construction

[0024] The invention provides an enhanced chemiluminescent detection kit, which includes liquid A and liquid B, wherein liquid A contains 0.1-10mM luminol, 1-10mM 3-(10'-phenothiazinyl)propane-1 - Sulfonate, and 0.5-3 mM 4-morpholinopyridine in 10-200 mM tris buffered saline solution; solution B is 1-10 mM urea peroxide solution.

[0025] Among them, the preparation method of A liquid is: adding formula amount of luminol, 3-(10'-phenothiazinyl) propane-1-sulfonate, tris buffer salt and 4-morpholinyl Pyridine, adjust the pH to 7.5-9.0 with hydrochloric acid, and constant volume with ultrapure water; the preparation method of B solution is to add the formula amount of carbamide peroxide, stir and dissolve with 1XPBS pH7.4 buffer solution, and constant volume.

[0026] The principle of the horseradish peroxidase luminol luminescence reaction catalyzed by the enhancer of the present invention is as follows: figure 1 shown. In the conventional ECL reaction, luminol, as the substr...

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Abstract

The invention discloses an enhanced chemiluminiscence detection kit. The enhanced chemiluminiscence detection kit comprises a solution A and a solution B, wherein the solution A is 10-200 mM of a trihydroxymethyl aminomethane buffer salt solution comprising 0.1-10 mM of luminol, 1-10 mM of 3-(10'-phenothiazinyl)propane-1-sulfonate, and 0.5-3 mM of 4-morpholinyl pyridine; and the solution B is 1-10mM of a urea peroxide solution. A novel enhancer and an electron transfer intermediate are adopted, the proportion of the components is adjusted, the more effective peroxide is matched, so that a more sensitive detection result is acquired, a target protein is detected from the pic level to the femtogram level; relatively high light intensity is kept within one hour after the reaction starts, thethermal acceleration experiment is conducted at the temperature of 37 DEG C for 2 weeks, but the product effect is not obviously affected, and the enhanced chemiluminiscence detection kit can be stored stably at the temperature of 4 DEG C for 1 year.

Description

technical field [0001] The invention relates to the field of biochemical experiment reagents, more particularly, to an enhanced chemiluminescence detection kit and a preparation method thereof. Background technique [0002] ECL (Enhance chemluminesence) is a luminescent reagent used in western blot. In western blot experiments, complex protein mixtures are separated by SDS-PAGE and transferred to solid-phase membranes (such as NC, PVDF) for immunological detection. primary antibody) or indirect (labeled secondary antibody) reaction; when ECL reagent is added, under the catalysis of horseradish peroxidase (HRP), luminol reacts with hydrogen peroxide to generate a peroxide, peroxide The substance is unstable and then decomposes to form a luminescent electronically excited intermediate. When the latter returns from the excited state to the ground state, it will produce fluorescence, and the reagent contains an enhancer to significantly enhance the luminescence. At present, th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N21/76
Inventor 肖扬季雨
Owner JIANGSU KEYGEN BIOTECH CORP LTD
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