Preparation method of nuclear matrix protein 22 enzyme-linked immunoassay kit and kit

An enzyme-linked immunoassay and nuclear matrix protein technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of high morbidity and postoperative recurrence rate, and achieve the effect of avoiding damage

Pending Publication Date: 2020-06-23
北京健平金星生物科技有限公司
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Problems solved by technology

Its incidence rate is relatively high, and with the change of lifestyle in recent years, it shows an increas

Method used

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preparation example Construction

[0037] The preparation method of a nuclear matrix protein 22 enzyme-linked immunoassay kit proposed by the present invention comprises the following steps:

[0038] The first step: preparing an enzyme-labeled conjugate of nuclear matrix protein 22 polyclonal antibody-labeled horseradish peroxidase; specifically,

[0039] Weigh 20-30 mg of horseradish peroxidase, dissolve it in 1%-1.5% glutaraldehyde solution and let it stand for 8-12 hours to make an enzyme solution;

[0040] After the enzyme solution was chromatographed on SephadexG-25, the enzyme solution was eluted with physiological saline, wherein the elution flow rate was controlled at 0.5-1.5ml / min, and the brown effluent was collected. If the volume is greater than 5ml, concentrate to 5ml with PEG; and stir in a container, specifically in a beaker;

[0041] Dilute 10-15 mg of nuclear matrix protein 22 to be labeled to 3-5 mg with physiological saline, and add it to the enzyme solution dropwise;

[0042] Add 0.2-0.5 m...

Embodiment 1

[0066] Step 1: Prepare the enzyme-labeled conjugate of nuclear matrix protein 22 polyclonal antibody-labeled horseradish peroxidase:

[0067] (1) take horseradish peroxidase 20mg and dissolve in 1% glutaraldehyde solution and let stand for 8 hours to make enzyme solution;

[0068] (2) After the enzyme solution was chromatographed on SephadexG-25, the enzyme solution was eluted with physiological saline, wherein the elution flow rate was controlled at 0.5 ml / min, and the brown effluent was collected. If the volume is greater than 5ml, concentrate to 5ml with PEG; and stir in a container, specifically in a beaker;

[0069] (3) Dilute 10-15 mg of nuclear matrix protein 22 to be labeled to 3 mg with physiological saline, and add it dropwise to the enzyme solution;

[0070] (4) Add 0.2 mg of carbonic acid buffer to the enzyme solution of nuclear matrix protein 22, and continue to stir for 2 hours; and add 0.2 mg of lysine after stirring, and wait for the first time to stand for 1 ...

Embodiment 2

[0079] Step 1: Prepare the enzyme-labeled conjugate of nuclear matrix protein 22 polyclonal antibody-labeled horseradish peroxidase:

[0080] (1) Take horseradish peroxidase 25 mg and dissolve it in 1%-1.5% glutaraldehyde solution and let it stand for 10 hours to make enzyme solution;

[0081] (2) After the enzyme solution was chromatographed on SephadexG-25, the enzyme solution was eluted with physiological saline, wherein the elution flow rate was controlled at 1 ml / min, and the brown effluent was collected. If the volume is greater than 5ml, concentrate to 5ml with PEG; and stir in a container, specifically in a beaker;

[0082] (3) Dilute 12.5 mg of nuclear matrix protein 22 to be labeled to 5 mg with physiological saline, and add it dropwise to the enzyme solution;

[0083] (4) Add 0.25 mg of carbonic acid buffer to the enzyme solution of nuclear matrix protein 22, and continue to stir for 3 hours; and add 0.25 mg of lysine after stirring, and wait for the first time to ...

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Abstract

The invention provides a preparation method of a nuclear matrix protein 22 enzyme-linked immunoassay kit and the kit. The method comprises the following steps: preparing an enzyme-labeled conjugate ofhorse radish peroxidase labeled by a nuclear matrix protein 22 polyclonal antibody; preparing a nuclear matrix protein 22 calibration product; preparing a solid phase coating plate; preparing a concentrated washing liquid; and preparing a stop solution, a substrate and a substrate buffer solution. The kit comprises a nuclear matrix protein 22 calibrator, a 96-well plate pre-coated with a nuclearmatrix protein 22 polyclonal antibody, an enzyme marker of the nuclear matrix protein 22 polyclonal antibody, a luminous substrate acting with an enzyme, a substrate buffer solution, a concentrated washing solution and a stop solution. According to the preparation method of the nuclear matrix protein 22 enzyme-linked immunoassay kit and the kit provided by the invention, early bladder cancer can be diagnosed, and damage to the body of a patient in the prior art is avoided.

Description

technical field [0001] The invention relates to the field of detection kits, in particular to a method for preparing a nuclear matrix protein 22 enzyme-linked immunological detection kit and the kit. Background technique [0002] Bladder cancer is one of the most common transitional cell carcinomas worldwide, accounting for approximately 90 percent of urothelial carcinomas. In the 2015 Cancer Annual Report, it ranked seventh among male cancers with 2.88%. Its incidence rate is relatively high, and with the change of lifestyle in recent years, it shows an increasing trend year by year. The main treatment method is surgery, but the postoperative recurrence rate is high. [0003] When cancer occurs, because the genetic material in the malignant cell nucleus cannot be distributed normally at the end of mitosis, the synthesis of nuclear mitotic apparatus protein increases sharply, and the level of NMP22 (nuclear matrix protein 22), which is a subunit of nuclear mitotic apparatus...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/577G01N33/543G01N33/535
CPCG01N33/57407G01N33/54306G01N33/535G01N33/577
Inventor 邹检平
Owner 北京健平金星生物科技有限公司
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