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P16 immune cell chemical labeling color-developing kit

A technology of immunocytochemistry and chromogenic reagents, which is applied in the field of p16 immunocytochemical labeling chromogenic kits, can solve the problems of poor positioning effect, poor labeling effect, and poor chromogenic effect, and achieve good positioning effect, obvious labeling, Good color effect

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 immunocytochemical labeling chromogenic kit to solve the problem in the above background technology that the current p16 immunocytochemical labeling chromogenic kit has poor positioning effect, poor color development effect and poor labeling effect during the experiment. bad question

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] A p16 immunocytochemical labeling color development kit, the detection kit includes the following reagents:

[0040] 0.5-1.0ml p16 immune cell standard concentration is 81pg / mL; 1.5-2.0ml standard diluent; 2-4ml horseradish peroxidase; 2-4ml sample diluent; 2-4ml chromogen A solution; 2-4ml developer B solution; 2-4ml stop solution; 20ml×20 times concentrated washing solution.

[0041] Further, p16 immune cell standard, standard diluent, horseradish peroxidase, sample diluent, chromogenic reagent A solution, chromogenic reagent B solution, stop solution and concentrated washing solution are all stored at 2-8°C. It is stored in the environment, and the storage time is more durable.

[0042] Specifically, after the kit is taken out of the refrigerated environment, it is placed at room temperature for 15-30 minutes before being used, and the test result is more accurate.

[0043] It is worth noting that the experimental use steps of the detection kit are as follows:

[...

Embodiment 2

[0057] As a second embodiment of the present invention, the samples are serum, plasma, urine, cell culture supernatant or tissue samples.

[0058] The specific operation of extracting the supernatant of the serum is as follows: the blood is naturally coagulated at room temperature for 10-20 minutes, then centrifuged for 15-20 minutes at a speed of 2000-3000 r / min, and the serum is collected.

[0059] The specific operation of extracting the supernatant from the plasma is as follows: add EDTA or sodium citrate as an anticoagulant to the plasma, mix for 10-20 minutes, centrifuge for 15-20 minutes at a speed of 2000-3000 r / min, and collect the serum.

[0060] The specific operation of extracting the supernatant from the urine is as follows: collect it with a sterile tube, centrifuge it for 15-20 minutes at a speed of 2000-3000 r / min, and collect the supernatant.

[0061] The specific operation of extracting the supernatant from the cell culture supernatant is as follows: when det...

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Abstract

The invention relates to the technical field of biotechnology, and specifically relates to a p16 immune cell chemical labeling color-developing kit. The detection kit comprises the following reagents:0.5-1.0 ml of a p16 immune cell standard substance with the concentration of 81 pg / mL, 1.5 to 2.0 ml of a standard substance diluent, 2-4 ml of horseradish peroxidase, 2-4 ml of a sample diluent, 2-4ml of a color-developing agent solution A, 2-4 ml of a color-developing agent solution B, 2-4 ml of a stop solution, and 20ml of 20 times of concentrated washing solution. The method comprises the steps of coating a microporous plate with a purified human p16 antibody to prepare a solid-phase antibody, adding p16 into micropores coated with a monoclonal antibody, combining with a horseradish peroxidase-labeled p16 antibody to form an antibody-antigen-enzyme-labeled antibody compound, thoroughly washing, and then adding a color-developing agent A substrate for color development. The color-developing agents A and B are converted into blue under the catalysis of horseradish peroxidase and converted into final yellow under the action of acid, the positioning effect and the color-developing effect are good, the labeling is more obvious, the color depth is in positive correlation with p16 in a sample, and the absorbance is measured by using a microplate reader at the wavelength of 450 nm.

Description

technical field [0001] The invention relates to the technical field of biotechnology, in particular to a p16 immunocytochemical labeling and color development kit. Background technique [0002] Human P16 protein is a protective protein in cells, which can prevent human cells from becoming cancerous and prevent excessive cell proliferation. The gene encoding human P16 protein is CDKN2A, which is located on the short arm of chromosome 9. P16 protein can bind to cyclin-dependent kinase 4 and inhibit its activity, so that cells stay in the G1 phase and cannot continue to proliferate. Therefore, P16 is a tumor suppressor protein. In cells, the expression of P16 protein is usually regulated by two other tumor suppressor proteins: P53 and RB, and is at a low level. [0003] The current p16 immunocytochemical labeling chromogenic kits on the market have poor positioning effect, poor chromogenic effect and poor labeling effect during the experiment. In view of this, we provide a ...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/535G01N21/31
CPCG01N33/54393G01N33/535G01N21/31
Inventor 黄子权
Owner 深圳市森盈生物科技有限公司
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