Staining agent for staining horse radish peroxidase and preparation method of staining agent, staining composition, application and marker enzyme staining kit

A technology of horseradish peroxidase and glucose oxidase, which is applied in the fields of analyzing materials through chemical reactions, material analysis and biological testing through observing the influence of chemical indicators, and can solve the problem of inconsistency in the resolution of HRP staining. High sensitivity, high resolution, reliable results, and suppression of false staining

Inactive Publication Date: 2017-05-10
舒斯云
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But this method has the following disadvantages: (1) the method is not sensitive, the dyeing of markers is not deep, it is difficult to dye a small amount of detection substances, and it is easy to produce false negative results, and it is easy to draw false conclusions that there is no object to be tested; (2) The specificity of this method is poor, and the part without markers may also be dyed yellow, and the non-specific substances are dyed together, which is easy to get a false positive result, resulting in a wrong result that is mistaken for the detected object; (3) the method The steps are complicated, the operation is troublesome, time-consuming and labor-intensive
In short, the resolution of HRP staining by this method is not high, and it is easy to cause misjudgment

Method used

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  • Staining agent for staining horse radish peroxidase and preparation method of staining agent, staining composition, application and marker enzyme staining kit
  • Staining agent for staining horse radish peroxidase and preparation method of staining agent, staining composition, application and marker enzyme staining kit
  • Staining agent for staining horse radish peroxidase and preparation method of staining agent, staining composition, application and marker enzyme staining kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Preparation of a marker enzyme staining kit:

[0050] Put 40 parts of diaminobenzidine, 10 parts of nickel ammonium sulfate, 90 parts of dextrose, 30 parts of ammonium chloride, 3 parts of glucose oxidase, 5 parts of acetic acid, 10 parts of sodium acetate, and 270 parts of distilled water into the 8 sealed bottles labeled 1-8, and then put into a carton to obtain a test kit.

[0051] A preparation of a staining agent for horseradish peroxidase staining:

[0052] First distilled water is divided into 90 parts of the first part of distilled water, 90 parts of the second part of distilled water and 90 parts of the third part of distilled water;

[0053] Then 40 parts of diaminobenzidine was mixed with 90 parts of the first part of distilled water to form the first mixture; 10 parts of nickel ammonium sulfate was mixed with 90 parts of the second part of distilled water to form the second mixture; 90 parts of dextrose, ammonium chloride 30 parts and 3 parts of glucose ox...

Embodiment 2

[0056] Preparation of a marker enzyme staining kit:

[0057] Put 60 parts of diaminobenzidine, 20 parts of nickel ammonium sulfate, 110 parts of dextrose, 50 parts of ammonium chloride, 8 parts of glucose oxidase, 15 parts of acetic acid, 30 parts of sodium acetate, and 330 parts of distilled water into the 8 sealed bottles labeled 1-8, and then put into a carton to obtain a test kit.

[0058] A preparation of a staining agent for horseradish peroxidase staining:

[0059] Firstly divide the distilled water into 110 parts of the first part of distilled water, 110 parts of the second part of distilled water and 110 parts of the third part of distilled water;

[0060] Then 60 parts of diaminobenzidine was mixed with 110 parts of the first part of distilled water to form the first mixture; 20 parts of nickel ammonium sulfate was mixed with 110 parts of the second part of distilled water to form the second mixture; 110 parts of dextrose, ammonium chloride 50 parts and 8 parts of ...

Embodiment 3

[0063] Preparation of a marker enzyme staining kit:

[0064] Put 50 parts of diaminobenzidine, 15 parts of nickel ammonium sulfate, 100 parts of dextrose, 40 parts of ammonium chloride, 5 parts of glucose oxidase, 10 parts of acetic acid, 20 parts of sodium acetate, and 300 parts of distilled water into the 8 sealed bottles labeled 1-8, and then put into a carton to obtain a test kit.

[0065] A preparation of a staining agent for horseradish peroxidase staining:

[0066] First divide the distilled water into 100 parts of the first part of distilled water, 100 parts of the second part of distilled water and 100 parts of the third part of distilled water;

[0067] Then 50 parts of diaminobenzidine was mixed with 100 parts of the first part of distilled water to form the first mixture; 15 parts of nickel ammonium sulfate was mixed with 100 parts of the second part of distilled water to form the second mixture; 100 parts of dextrose, ammonium chloride 40 parts and 5 parts of gl...

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Abstract

The invention relates to the technical field of biology and provides a staining agent for staining horse radish peroxidase and a preparation method of the staining agent, a staining composition, an application and a marker enzyme staining kit. The staining composition comprises the following reagents: diaminobenzidine, nickel ammonium sulfate, glucose, ammonium chloride, glucose oxidase, acetic acid and sodium acetate. The staining composition can be applied to preparation of the marker enzyme staining kit. The kit comprises the staining composition. The staining agent is prepared from the staining composition by the processes of preparing a diaminobenzidine water solution from diaminobenzidine as a first mixture; preparing a nickel ammonium sulfate water solution from nickel ammonium sulfate as a second mixture; mixing the glucose, the ammonium chloride and the glucose oxidase to form a third mixture; preparing a sodium acetate water solution from the acetic acid and the sodium acetate as a fourth mixture; and mixing the first mixture and the second mixture, then mixing with the third mixture and finally mixing with the fourth mixture. The preparation method is simple in procedure.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a sensitive staining agent for horseradish peroxidase staining, a preparation method, a staining combination, an application and a labeling enzyme staining kit. Background technique [0002] Horseradish peroxidase (HRP) is the most commonly used qualitative and quantitative labeling enzyme in biological, medical and pharmaceutical experiments and detection. It can be used to label proteins, nucleic acids, viruses, tumor factors, drugs, poisons, cells Factors, antibodies, antigens, receptors, and specific components in blood, cerebrospinal fluid, and tissues, etc. HRP itself has no color, and the nature and content of the detected substance can be qualitatively or quantitatively developed only by dyeing method. [0003] The most commonly used HRP staining method is diaminobenzidine staining. But this method has the following disadvantages: (1) the method is not sensitive, the dyeing...

Claims

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Application Information

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IPC IPC(8): G01N33/535G01N33/52G01N21/78
CPCG01N21/78G01N33/52G01N33/535
Inventor 舒斯云
Owner 舒斯云
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