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Detecting method of pig metal sulfur protein

A metallothionein and detection method technology, applied in the field of qualitative and quantitative detection of porcine metallothionein, can solve the problems of radioactive contamination, unsatisfactory repeatability, stability and sensitivity, and low sensitivity, and achieve stability and repeatability. Good, avoid radioactive contamination, high sensitivity

Inactive Publication Date: 2005-05-25
HUNAN AGRICULTURAL UNIV
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AI Technical Summary

Problems solved by technology

[0004] At present, the cadmium-hemoglobin saturation method is a classic method recognized by the academic community for the detection of metallothionein, but this method cannot be used for the detection of Bi, Hg, Ag, and Cu-bound metallothionein, and its sensitivity is not high, so it cannot be used for detection The content of metallothionein in various tissues and organs of animals under normal physiological state
The use of radioimmunoassay requires high experimental conditions and radioactive contamination, so it cannot be widely promoted
Some scholars have also begun to try other types of enzyme-linked immunosorbent assay (ELISA), but the repeatability, stability and sensitivity of the results are not satisfactory. The need for further research

Method used

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Examples

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Embodiment Construction

[0022] Describe in detail below in conjunction with specific embodiment:

[0023] 1. Detection

[0024] ①Preparation of porcine plasma samples: 9-11ml of blood from the hepatic portal vein was extracted through the hepatic portal vein vascular catheter, and 10% EDTA-Na was used as an anticoagulant, centrifuged for 20min, 3000rpm, 4°C, and the plasma was subpackaged and stored at -70°C. to be tested;

[0025] Preparation of pig tissue and organ samples: After weighing pig liver, kidney and muscle, add Tris-HCl buffer (0.1mol / l, pH8.6) containing 0.25mol / l sucrose in 1 / 4 amount, homogenate, Centrifuge for 20min (8000rpm, 4°C), take the supernatant and put it in -70°C for storage until testing;

[0026] ②Coating: Dilute the standard MT with coating buffer (pH=9.6) to a concentration of 6.25 μg / ml, add 100 μl / well of coating solution to the microplate plate, seal it with a parafilm, and put it in 4 overnight at ℃ (16-24hr), take it out, add 250μl of washing solution to each wel...

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Abstract

The present invention disclosed a detection method of swine metallothionein, relates to detection method of animal metallothionein, it relates to a qualitative and quantitative detection method of swine metallothionein. Said invention uses electrophoretic pure mouse anti-swine metallothionein IgG as first antibody and uses goat anti-mouse enzyme-linked antibody as second antibody, and adopts solid-phase antigen indirect competitive type enzyme-linked immunosorbent assay method to determine the content of swine metallothionein. Said invention also provides the concrete steps of said method.

Description

technical field [0001] The invention relates to a detection method for animal metallothionein, in particular to a qualitative and quantitative detection method for pig metallothionein. Background technique [0002] Metallothionein (MT) has a strong effect of eliminating free radicals, and can be used alone as a nutritional supplement or in combination with glutathione (GSH), vitamin C (VC), vitamin E (VE), etc., which can effectively inhibit Lipid peroxidation damage, cell protection, lower blood viscosity, improve blood circulation and enhance the body's immunity and other biological functions. Metallothionein can also be used as an antioxidant to prevent oxidative deterioration of food or feed. In addition, metallothionein can combine with many metal ions, participate in the regulation of metal metabolism in the body, balance the homeostasis of metal ions in the body, participate in the operation and excretion of metal ions, and play an important role in mineral nutrition...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/543
Inventor 张彬吴力专李丽立
Owner HUNAN AGRICULTURAL UNIV
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