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Method for deactivating endogenous peroxydase and application thereof

A peroxidase, endogenous technology, used in biochemical equipment and methods, enzymes, microorganisms, etc., can solve problems such as non-essential, toxic effects, dyeing effects, etc., to reduce influencing factors, simple operation, and operation steps Streamlined effect

Inactive Publication Date: 2011-05-18
PEKING UNIV THIRD HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that metal ions are not only unnecessary for antigen retrieval, but also their toxic effects may affect the final staining effect

Method used

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  • Method for deactivating endogenous peroxydase and application thereof
  • Method for deactivating endogenous peroxydase and application thereof
  • Method for deactivating endogenous peroxydase and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1, block endogenous peroxidase by microwave heating

[0038] One, each experimental method used in the present embodiment:

[0039] The experimental methods used in this example are shown in Table 1.

[0040] Table 1. Different endogenous peroxidase blocking procedures

[0041] method

serial number

Method steps

M1

Dewaxing and rehydration→DAB color development→sealing, microscope observation

M2

Dewaxing and rehydration→microwave high temperature heat treatment→DAB color development→sealing, microscope observation

M3

Dewaxing rehydration → 3% H 2 o 2 → Microwave high temperature heat treatment → DAB color development → sealing,

microscope observation

M4

Dewaxing rehydration → 0.3% H 2 o 2 → Microwave high temperature heat treatment → DAB color development → sealing,

microscope observation

M5

Dewaxing and rehydration→microwave high temperature heat treatme...

Embodiment 2

[0109] Example 2. Application of Microwave Heating to Block Endogenous Peroxidase Activity in Immunohistochemistry

[0110] In this example, two methods were used to observe the expression of Bax antigen and Bcl-2 antigen in tissues. The first method is the traditional immunohistochemical method, and the second method is to block endogenous peroxidase by microwave heating According to the immunohistochemical method, by comparing the antigen detection effect and the immunohistochemical staining effect, it can be seen whether the immunohistochemical method using microwave heating to block endogenous peroxidase of the present invention is effective.

[0111] 1. Materials and reagents used in the experiment

[0112] Materials and sources: In this example, human gastric cancer tissues and the above-mentioned colon and small intestine tissues were used as experimental materials. Sources of materials: Colon and small intestine tissue samples were from the same source as above, and g...

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Abstract

The invention discloses a method for deactivating endogenous peroxydase in the immunohistochemistry, which deactivates the endogenous peroxydase by using a microwave treated tissue slice, wherein the frequency is 2.45GHz, the power is 650-800w, and the treatment time is 10-15min. Proved by experiments to various tissues, the method can deactivate the endogenous peroxydase, can play roles of antigen and restoration and has simple operation and time and labor saving, therefore, the application of the method in the immunohistochemistry can merge the antigen and the restoration into the confinement of the endogenous peroxydase, the step of deactivating the endogenous peroxydase by using diluted perhydrol confining liquid can be omitted to lead the operation of the immunohistochemistry to be simpler, and the influence factor is reduced further.

Description

technical field [0001] The invention relates to a method for inactivating endogenous peroxidase and its application. Background technique [0002] Immunohistochemical technology has been widely used in clinical and scientific research, and can provide more important information for the localization of proteins in tissues and related functional studies. There are many factors affecting the staining effect, including endogenous peroxidase treatment, microwave antigen retrieval process, antibody, tissue fixative, antigen location, and DAB color development time, etc. Whether endogenous peroxidase can be successfully blocked is currently considered to be one of the key steps affecting the effect of immunohistochemical staining, because if the endogenous peroxidase activity in the tissue cannot be completely blocked, it will cause immunohistochemical staining. False positive staining greatly interferes with the judgment of the results. The use of dilute hydrogen peroxide blocki...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/99C12N13/00
Inventor 高春王爱英韩亚京
Owner PEKING UNIV THIRD HOSPITAL