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dab chromogenic kit for immunohistochemical detection and its application

A chromogenic reagent and immunohistochemical technology, which can be used in biological testing, analysis through chemical reaction of materials, material analysis through observation of the influence of chemical indicators, etc., can solve complex components and color development time Problems such as prolongation and over-fast response can achieve the effects of improving stability and sensitivity, increasing strength and sensitivity, and reducing non-specific binding

Active Publication Date: 2022-06-24
中关村科技租赁股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The DAB kits used in the existing immunohistochemical method have various components, and most of them have the following problems: (1) The amount of hydrogen peroxide will affect the intensity and stability of immunohistochemical color development
If the hydrogen peroxide content is too low, the color development will be too weak or the color development time will be prolonged; if the hydrogen peroxide concentration exceeds the appropriate concentration, the reaction will be too fast and the background will be deepened
Because hydrogen peroxide will slowly decompose into water and oxygen in general environment, the stability of hydrogen peroxide in conventional DAB substrate buffer is poor and the validity period is short; (2) some DAB chromogenic kits are used to protect the To oxidize hydrogen and delay the oxidation of DAB, various protective agents or stabilizers will be added, making its components complex and cumbersome; (3) the detection sensitivity of conventional DAB kits to low-abundance antigens is low; (4) conventional In the DAB kit, after the chromogen and the substrate buffer are mixed, it needs to be prepared and used immediately, and cannot be stored for a long time

Method used

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  • dab chromogenic kit for immunohistochemical detection and its application
  • dab chromogenic kit for immunohistochemical detection and its application
  • dab chromogenic kit for immunohistochemical detection and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Preparation of DAB buffer A#1-A#4

[0052] This example provides 4 kinds of DAB buffers A#1-A#4, all including imidazole, 4-nonylphenyl-polyethylene glycol, benzalkonium chloride, hydrogen peroxide, EDTA and sodium pyrophosphate, the difference is only The amount of hydrogen peroxide is different, and the specific components are shown in Table 1.

[0053] Table 1 DAB buffer A#1-A#4 components

[0054]

[0055] The preparation method is as follows:

[0056] Accurately weigh or measure imidazole, 4-nonylphenyl-polyethylene glycol, benzalkonium chloride, hydrogen peroxide, EDTA and sodium pyrophosphate in a clean and sterilized container, add 200 mL of purified water and mix thoroughly. , adjust the pH to 7.6 with hydrochloric acid, and add purified water to the volume to 250mL. After thorough mixing, filter with a 0.22 μm filter and save for later use.

Embodiment 2

[0057] Example 2 Preparation of DAB buffer A#5-A#11

[0058] This example provides seven DAB buffers, A#5-A#11, all including imidazole and 4-nonylphenyl-polyethylene glycol, but also benzalkonium chloride, hydrogen peroxide, EDTA, and / or pyrophosphate Sodium, the specific components are shown in Table 2.

[0059] Table 2 DAB buffer A#5-A#11 components

[0060]

[0061]

[0062] The preparation method is as follows:

[0063] Accurately weigh or measure one or two or three of imidazole, 4-nonylphenyl-polyethylene glycol, and benzalkonium chloride, hydrogen peroxide, EDTA and sodium pyrophosphate in a clean sterilized container , add 200 mL of purified water, stir and mix well, adjust the pH to 7.6 with hydrochloric acid, and add purified water to make the volume to 250 mL. After thorough mixing, filter with a 0.22 μm filter and save for later use.

Embodiment 3

[0064] Example 3 Preparation of DAB buffer A#12-A#15

[0065] This example provides 4 kinds of DAB buffers A#12-A#15, all including imidazole, 4-nonylphenyl-polyethylene glycol, benzalkonium chloride, hydrogen peroxide, EDTA and sodium pyrophosphate, the difference is The pH values ​​are different, and the specific components are shown in Table 3.

[0066] Table 3 DAB buffer A#12-A#15 components

[0067]

[0068] The preparation method is as follows:

[0069] Accurately weigh or measure imidazole, 4-nonylphenyl-polyethylene glycol, benzalkonium chloride, hydrogen peroxide, EDTA and sodium pyrophosphate in a clean and sterilized container, add 200 mL of purified water and mix thoroughly. , adjusted to the desired pH with hydrochloric acid, and then added purified water to make up to 250 mL. After thorough mixing, filter with a 0.22 μm filter and save for later use.

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Abstract

The invention discloses a DAB chromogenic kit for immunohistochemical detection, belonging to the technical field of immunohistochemical detection. The DAB chromogenic kit includes DAB buffer and DAB chromogen, and the DAB buffer includes imidazole, 4-nonylphenyl-polyethylene glycol, benzalkonium chloride, hydrogen peroxide, EDTA solution and pyrophosphoric acid Sodium; the DAB chromogen includes: 1,2-propylene glycol and 3,3-diaminobenzidine tetrachlorosalt hydrate, further, the kit also includes a DAB enhancer, and the DAB enhancer includes copper sulfate, Sodium oxide, potassium oxide, sucrose and benzalkonium chloride. The invention further discloses the application of the DAB chromogenic kit. Using the DAB chromogenic kit of the present invention for immunohistochemical detection can significantly improve the staining intensity and sensitivity, and the DAB chromogenic kit of the present invention has simple components, is easy to prepare, and has excellent stability.

Description

technical field [0001] The invention belongs to the technical field of immunohistochemical detection, in particular to a DAB color development kit for immunohistochemical detection and application thereof. Background technique [0002] Immunohistochemistry (abbreviated as immunohistochemistry) is a detection method that uses the principle of specific binding of antigens and antibodies to color the substrate of labeled antibodies through chemical reaction to determine whether there is a target antigen in tissue and its expression. It combines the specificity of the immune response and the visibility of histochemistry, and uses the imaging and magnification of microscopes (including fluorescence microscopes and electron microscopes) to detect various antigenic substances at the cellular and subcellular levels, and to locate and magnify them. Qualitative and quantitative research. The continuous development of immunohistochemical technology has brought great help to clinical p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/78G01N33/58G01N33/53
CPCG01N21/78G01N33/581G01N33/53G01N2021/775
Inventor 陶娜娜章月凯潘丽
Owner 中关村科技租赁股份有限公司
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