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Human S100 protein detection reagent and preparation method thereof

A protein detection and reagent technology, applied in the field of laboratory medicine, can solve the problems of difficulty in ensuring the stability and repeatability of test results, inconvenient for routine laboratory development, and increasing the burden on patients, so as to improve the efficiency of detection work and the cost is not high. , the effect of easy operation

Active Publication Date: 2014-02-26
江苏美德医药技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is divided into two types, one is the direct labeling method, which belongs to the instant luminous type, it is difficult to guarantee the stability and repeatability of the test results, and it requires special testing instruments
Not convenient for routine laboratory development
The second biotin-avidin-labeled electrochemiluminescence immunoassay method needs to be equipped with an expensive automatic electrochemiluminescence detector, which cannot be carried out in conventional laboratories, and it brings unnecessary costs to patients and increases the burden on patients

Method used

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  • Human S100 protein detection reagent and preparation method thereof
  • Human S100 protein detection reagent and preparation method thereof
  • Human S100 protein detection reagent and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] 1. The R1 reagent: HEPES buffer

[0060]

[0061] After dissolving the above raw materials with 800ml purified water, add purified water to make the volume to 1L, and then adjust the pH to 8.0 with hydrochloric acid or sodium hydroxide.

[0062] 2. The R2 reagent:

[0063] Step 1: Dilute 300nm carboxyl latex microspheres with 50mmol / L, PH6.0 MES buffer to a 0.01mg / ml suspension; according to the ratio of 1ml carboxyl latex microsphere suspension to 20mg EDAC and 40mg NHS , Add EDAC and NHS and mix them immediately, then react the mixture at room temperature for 15-30 minutes, stirring constantly;

[0064] Step 2: Wash the carboxyl latex microspheres with 50mmol / L, PH6.0 MES buffer or purified water to remove unreacted NHS and EDAC, and suspend the latex microspheres in purified water to obtain a concentration of 0.01mg / ml Suspension of carboxyl latex microspheres;

[0065] Step 3: Dissolve the mouse anti-human S100 antibody in a 50mmol / L, PH8.5 TAP salt buffer solution so that ...

Embodiment 2

[0075] 1. The R1 reagent: Tris buffer

[0076]

[0077]

[0078] After dissolving the above raw materials with 800ml of purified water, add purified water to make the volume up to 1L, and then adjust the pH to 7.4 with hydrochloric acid or sodium hydroxide.

[0079] 2. The R2 reagent:

[0080] Step 1: Take 100nm carboxyl latex microspheres and dilute with 50mmol / L, PH6.0 MES buffer into 0.01mg / ml suspension; according to the ratio of 1ml carboxyl latex microsphere suspension to 20mg EDAC and 40mg NHS , Add EDAC and NHS and mix them immediately, then react the mixture at room temperature for 15-30 minutes, stirring constantly;

[0081] Step 2: Wash the carboxyl latex microspheres with 50mmol / L, PH6.0 MES buffer or purified water to remove unreacted NHS and EDAC, and suspend the latex microspheres in purified water to obtain a concentration of 0.01mg / ml Suspension of carboxyl latex microspheres;

[0082] Step 3: Dissolve the mouse anti-human S100 antibody in a 50mmol / L, PH8.5 TAP salt b...

Embodiment 3

[0093] 1. The R1 reagent: Glycine buffer

[0094]

[0095] After dissolving the above raw materials with 800ml purified water, add purified water to make the volume to 1L, and then adjust the pH to 8.0 with hydrochloric acid or sodium hydroxide. Among them, Thesit is a polyoxyethylene lauryl ether, which is of the same kind as Tween-20.

[0096] 2. The R2 reagent:

[0097] Step 1: Dilute 200nm carboxyl latex microspheres with 50mmol / L, PH6.0 MES buffer to a 0.01mg / ml suspension; according to the ratio of 1ml carboxyl latex microsphere suspension to 20mg EDAC and 40mg NHS , Add EDAC and NHS and mix them immediately, then react the mixture at room temperature for 15-30 minutes, stirring constantly;

[0098] Step 2: Wash the carboxyl latex microspheres with 50mmol / L, PH6.0 MES buffer or purified water to remove unreacted NHS and EDAC, and suspend the latex microspheres in purified water to obtain a concentration of 0.01mg / ml Suspension of carboxyl latex microspheres;

[0099] Step 3: Di...

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Abstract

The invention relates to a human S100 protein detection reagent and a preparation method thereof, and particularly relates to a reagent for detecting a human S100 protein by virtue of latex-enhanced immunoturbidimetry and a preparation method thereof. The reagent comprises a reagent R1, a reagent R2 and a standard product and is characterized in that the reagent R1 is a modified buffer solution with the pH value of 7.0-8.0; the reagent R2 is an anti-human S100 protein antibody latex reagent; the standard product is recombinant protein containing quantitative S100 protein or natural S100 protein standard liquid extracted from human serum. Compared with the prior art, the reagent provided by the invention has the characteristics of (1) high accuracy (the correlation R2 with the chemiluminescence immunoassay is 0.9956-0.9995) and (2) fast detection (only 10 minutes are needed from the test start to the result acquisition). Large-batch sample detection can be performed on a conventional biochemical analyzer, and the detection efficiency is greatly improved. The preparation method of the reagent is simple to operate, the raw materials are easily available, the cost is not high, and the method is suitable for various medical research institutions and conventional laboratories.

Description

Technical field [0001] The invention relates to a reagent for detecting human S100 protein and a preparation method, in particular to a reagent and preparation method for detecting human S100 protein by adopting latex-enhanced immunoturbidimetric method, which can be applied in the field of laboratory medicine. Background technique [0002] Human central nervous system protein (S-100 protein) is mainly concentrated in the astrocytes of the central nervous system and corresponding tumor cells. The S-100 molecule is composed of α and β subunits with a molecular weight of 21,000 and a biological half-life of 22 hours. . The S-100 protein in cerebrospinal fluid is related to age and increases with age, but the concentration of S-100 protein in plasma has no exact relationship with age and gender. When the central nervous system cells are injured, S100 protein leaks from the cytosol into the cerebrospinal fluid, and then enters the blood through the damaged brain blood barrier. Ther...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/566
CPCG01N33/531G01N33/538G01N33/6896G01N33/96G01N2496/00G01N2800/28
Inventor 陆上苏
Owner 江苏美德医药技术有限公司