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Anti-androgen effect detection method

A detection method, an anti-androgen technology, applied in the field of detection of anti-androgen effects, can solve the problems of high cost of time and money, large error in test results, and many interference factors, and achieve simplified operation, high continuous stability, and reduced cost effect

Active Publication Date: 2018-12-21
INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, in vivo detection is a commonly used detection method, but the experiment is cumbersome, and the cost of time and money is high.
However, the traditional in vitro detection method has high cost, many interference factors, and large error in test results.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0095] 1. Determination of cytotoxic activity by MTT colorimetry:

[0096] 1. Adjust the cell density with L-15 medium containing 10% fetal bovine serum, 1×10 per hole in 100ul 4 Cells were seeded into 96-well culture plates at 37°C, 5% CO 2 Incubate for 24 h under conditions.

[0097] 2. Remove the culture medium, add 100ul of pesticide-containing medium without fetal bovine serum, and 0.1% DMSO as a blank control group.

[0098] 3. Incubate with pesticides for 24 hours, and remove the drug-containing medium.

[0099] 4. Add 110ul 10% MTT solution, prepare it with serum-free medium, and incubate for 3h.

[0100] 5. Detect the absorbance value of each well at 490nm with a microplate reader.

[0101] Cell viability was calculated using the formula:

[0102] Cell Viability / %=(OD sample-OD blank) / (OD control-OD blank)×100.

[0103] 2. Determination method of anti-androgen effect of pesticides:

[0104] 1. Using the MDA-kb2 breast cancer cell line, adjust the cell density w...

Embodiment 2

[0111] 1. Determination of cytotoxic activity by MTT colorimetry:

[0112] 1. Adjust the cell density with L-15 medium containing 9% fetal bovine serum, 0.8×10 per hole in 100ul 4 Cells were seeded into 96-well culture plates at 37°C, 5% CO 2 Incubate for 23 h under conditions.

[0113] 2. Remove the culture medium, add 100ul of pesticide-containing medium without fetal bovine serum, and 0.1% DMSO as a blank control group.

[0114] 3. Incubate with pesticides for 24 hours, and remove the drug-containing medium.

[0115] 4. Add 110ul 10% MTT solution, prepare it with serum-free medium, and incubate for 3h.

[0116] 5. Detect the absorbance value of each well at 490nm with a microplate reader.

[0117] Cell viability was calculated using the formula:

[0118] Cell Viability / %=(OD sample-OD blank) / (OD control-OD blank)×100.

[0119] 2. Determination method of anti-androgen effect of pesticides:

[0120] 1. Using the MDA-kb2 breast cancer cell line, adjust the cell density ...

Embodiment 3

[0127] 1. Determination of cytotoxic activity by MTT colorimetry:

[0128] 1. Adjust the cell density with L-15 medium containing 11% fetal bovine serum, 1.2×10 per hole in 100ul 4 Cells were seeded into 96-well culture plates at 37°C, 5% CO 2 Incubate for 26 h under conditions.

[0129] 2. Remove the culture medium, add 100ul of pesticide-containing medium without fetal bovine serum, and 0.1% DMSO as a blank control group.

[0130] 3. Incubate with pesticides for 24 hours, and remove the drug-containing medium.

[0131] 4. Add 110ul 10% MTT solution, prepare it with serum-free medium, and incubate for 3h.

[0132] 5. Detect the absorbance value of each well at 490nm with a microplate reader.

[0133] Cell viability was calculated using the formula:

[0134] Cell Viability / %=(OD sample-OD blank) / (OD control-OD blank)×100.

[0135] 1. Using the MDA-kb2 breast cancer cell line, adjust the cell density with L-15 medium containing 11% fetal bovine serum to contain 1.2×10 4 ...

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PUM

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Abstract

The invention relates to the field of anti-androgen effect detection, in particular to an anti-androgen effect detection method. The detection method comprises the steps that the anti-androgen and ligand substance in a to-be-detected sample compete for an androgen receptor in the cells, and the effect value of the anti-androgen can be determined by measuring the amount of the ligand substance combined with the receptor, and the effect value is evaluated by taking the cell toxicity activity result of the to-be-detected sample on the cells as a reference factor, wherein the ligand substance is aligand of the androgen receptor; and the competition is combined in a serum-free reaction system. By adoption of to the method, the influences of hormone and growth factors in serum and the cell toxicity activity of the to-be-detected product on fluorescence intensity detection are eliminated, the cost can be lowered, the experiment error can be reduced, and the experiment process can be simplified.

Description

technical field [0001] The invention relates to the field of detection of anti-androgen effect, in particular to a detection method of anti-androgen effect. Background technique [0002] In recent years, the impact of environmental endocrine disruptors (Environmental endocrine disrupters, EEDs) on the endocrine system of wild animals and humans, especially on reproductive endocrine function has attracted great attention, among which the impact on male reproductive health is the focus. Malformations of the reproductive system, reduced or lost fertility, reduced hatchability of bird and amphibian eggs, increased embryo mortality, larval survival, and Reproductive degradation phenomena such as reduction and obvious reduction in population number. Environmental antiandrogen (EA) is an environmental endocrine disruptor. Like environmental estrogen, it can cause abnormal development and function of the male reproductive system. Now it is an increase in the incidence of urogenital...

Claims

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Application Information

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IPC IPC(8): G01N33/74G01N33/68
CPCG01N33/6803G01N33/74G01N2333/723G01N2333/47
Inventor 钱永忠陈晨马朦朦王天彩李耘许彦阳
Owner INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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