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Identification method for blood serum proteome of broiler chickens

A proteomics and identification method technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of low specificity, limited application, and increased cost, and achieve high-sensitivity detection, elimination of individual differences, and low cost Effect

Inactive Publication Date: 2017-05-31
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

The protein precipitation method uses acetonitrile, ethanol, acetone, etc. to precipitate high-abundance proteins in serum to varying degrees, but the specificity of this method is not high; affinity technology uses the interaction between proteins and their ligands or antibodies to specifically bind High-abundance target proteins such as albumin and IgG have high specificity, but the cost of this method increases, and it is worth noting that most of the existing commercial products are developed for human and mouse serum, which limits its application. limit
[0004] At present, there is no relevant report on the identification method of broiler serum proteome

Method used

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  • Identification method for blood serum proteome of broiler chickens
  • Identification method for blood serum proteome of broiler chickens
  • Identification method for blood serum proteome of broiler chickens

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Experimental program
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Embodiment Construction

[0028] Such as figure 1 Shown, the concrete realization process of the inventive method is as follows:

[0029] (1) Serum sample collection: collect the blood of 5 normal broilers, put it in a negative pressure tube without heparin, and set the negative pressure tube on ice for 15-20min, at a temperature of 4-8°C and a centrifugal force of 1600-1800g The supernatant was taken after centrifuging for 15-20 minutes under the conditions, and the obtained serum samples were quantified for protein and mixed with equal masses.

[0030](2) SDS-PAGE electrophoresis: ① Take 20 μg of protein and carry out SDS-PAGE electrophoresis with a mass fraction of 12%. Electrophoresis conditions: after 30 minutes of electrophoresis at 80V, electrophoresis at 110V until bromophenol blue reaches the front edge of the gel; ② Stain the gel obtained in step ① with Coomassie Brilliant Blue staining method: place the gel in 0.1% R-250 staining solution, shake and stain on a shaker at 25-40°C for 2 hours,...

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Abstract

The invention provides an identification method for blood serum proteome of broiler chickens. The identification method comprises the following steps: (1) collecting a blood serum sample; (2) carrying out SDS-PAGE (Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis); (3) carrying out in-gel enzymolysis on protein gel; (4) analyzing protein by nano-HPLC-MS / MS (nano-High Performance Liquid Chromatography-Mass Spectrometry / Mass Spectrometry); searching a protein data base through Protein Pilot software to identify obtained data. By adopting the method provided by the invention, the quantity and types of blood serum protein expression of the broiler chickens can be rapidly found at high throughput, and a method and a concept are provided for development of research on nutrients of the broiler chickens and immune functions of the broiler chickens.

Description

technical field [0001] The invention relates to the fields of analytical chemistry and life sciences, in particular to an identification method for broiler serum proteome. Background technique [0002] Serum has the functions of maintaining the normal viscosity, pH, and osmotic pressure of blood. The protein source in serum is related to almost all cells, tissues, and organs, so it can directly reflect the pathological and physiological state of the body. It is the key to disease diagnosis and biomarker discovery. important specimen. Compared with tissue proteins and cellular proteins, serum proteins have a wide variety and different properties, and there are also a large number of protein splicing bodies, post-translational modifications, and degradation fragments. In addition, the abundance of serum proteins varies greatly (the protein concentration range spans 10 orders of magnitude), and high-abundance proteins account for more than 99% of the total protein, which serio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/06
CPCG01N30/88G01N30/06G01N2030/8831
Inventor 唐湘方熊嫣张宏福孟庆石高杰冯潇慧
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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