P16 immunohistochemical detection kit

A detection kit and immunohistochemical technology, applied in the field of biotechnology, can solve the problems of inaccurate qualitative positioning, long incubation time, inconvenient operation, etc., and achieve the effect of convenient operation, short incubation time and clear background

Pending Publication Date: 2019-11-26
深圳市森盈生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a p16 immunohistochemical detection kit to solve the problem of low sensitivity, poor specificity, inaccurate qualitative positioning, blurred background, and inconvenient operation of the current p16 immunohistochemical detection kit proposed in the above background technology. Problem with too long incubation time

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] A p16 immunohistochemical detection kit, including the following reagents in the detection kit:

[0045] Primary antibody: lyophilized p16 antibody;

[0046] Colorless reagent: endogenous peroxidase blocker 10-15ml;

[0047] Reagent A: Immunohistochemical blocking solution;

[0048] Reagent B: Biotin-labeled secondary antibody working solution;

[0049] Reagent C: horseradish enzyme-labeled streptavidin working solution;

[0050] Brown chromogenic solution D solution 1-2ml;

[0051] White chromogenic solution E solution 20-25ml;

[0052] 0.1M PBS buffer;

[0053] Sodium citrate antigen restoration solution;

[0054] Diaminobenzamine DAB staining solution.

[0055] PBS (phosphate buffered solution) is a phosphate buffer solution, which can provide a relatively stable ionic environment and pH buffering capacity. It is a buffered salt solution often used in biology and is used for molecular cloning and cell culture. The pH is 7.2-7.4. Isotonic with human blood, the...

Embodiment 2

[0074] As the second embodiment of the present invention, the primary antibody is diluted using the following two methods:

[0075] (1) Dilute the lyophilized p16 antibody with 0.1M PBS buffer, divide the diluted antibody into 5-10 vials, 20ulx5 vials or 10ulx10 vials, and store at -20°C;

[0076] (2) Dilute the lyophilized p16 antibody with 0.1M PBS buffer solution to 50ul to form a stock solution, add 50% glycerol, and store at -20°C.

[0077] Specifically, the preparation experiment of the DAB chromogenic solution specifically includes the following steps: use 1ml of the white chromogenic solution E solution as the DAB substrate solution, and then add 1 drop of 50ul brown chromogenic solution D solution as the DAB concentrated solution, mix well, Prepare the DAB chromogenic solution. This solution must be prepared immediately for use. After preparation, it should be stored away from light. It should be used within 6 hours. The remaining liquid should be discarded. The color...

Embodiment 3

[0079] The repair methods of sodium citrate antigen repair solution include boiling water bath repair, microwave repair or high pressure repair. Sodium citrate antigen repair solution can be used for paraffin sections, frozen sections and other samples fixed with paraformaldehyde, formaldehyde or other aldehyde reagents. Antigen retrieval can effectively remove the cross-linking between proteins caused by aldehyde fixation reagents, and fully expose the antigenic epitopes in samples such as paraffin sections, thereby greatly improving the effect of immunostaining.

[0080] Specifically, for boiling water bath repair, place the beaker containing the repair solution and glass slides in a boiling water bath environment, keep the external boiling state for 15 minutes, and cool naturally to room temperature.

[0081] For microwave repair, put the beaker containing the repair solution and glass slides in a microwave oven, set high heat for 5 minutes, stop fire for 3 minutes, medium h...

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PUM

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Abstract

The invention relates to the technical field of biotechnology, and specifically relates to a p16 immunohistochemical detection kit. The p16 immunohistochemical detection kit comprises a primary antibody, namely, a freeze-dried powder p16 antibody, a colorless reagent, namely, 10-15ml of an endogenous peroxidase blocking agent, a reagent A, namely, an immunohistochemical confining liquid, a reagentB, namely, a biotin labeled secondary antibody working solution, a reagent C, namely, a horseradish peroxidase labeled streptavidin working solution, 1-2ml of a brown color developing solution D, 20-25ml of a white color developing solution E, 0.1M of a PBS buffer solution, a sodium citrate antigen retrieval solution and a diamino-benzidine DAB staining solution. The immunohistochemical detectionreagent produced by using a streptavidin-biotin signal amplification technology has the advantages of high sensitivity, strong specificity, accurate qualitative positioning, clear background, low-power scene staining, direct dropwise addition, convenient operation and short incubation time, and is suitable for frozen sections, paraffin sections and fixed cell smears; and the kit can be used for detecting specific antigens in cells and tissues.

Description

technical field [0001] The invention relates to the technical field of biotechnology, in particular to a p16 immunohistochemical detection kit. Background technique [0002] Immunohistochemistry is the use of the principle of specific binding of antigens and antibodies, through chemical reactions to make the chromogenic reagents (fluorescein, enzymes, metal ions, isotopes) of labeled antibodies develop color to determine the antigens (polypeptides and proteins) in tissue cells. It conducts localized, qualitative and quantitative research. At present, immunohistochemistry has been widely used in clinical pathological diagnosis, especially in the auxiliary diagnosis and differential diagnosis of tumors with targeted antibodies, which has become an indispensable and important means for clinical pathological diagnosis. [0003] The current p16 immunohistochemical detection kits on the market have low sensitivity, poor specificity, inaccurate qualitative positioning, blurred bac...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/532G01N1/30
CPCG01N33/54393G01N33/532G01N1/30G01N2001/302
Inventor 黄子权
Owner 深圳市森盈生物科技有限公司
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