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Using method of iTRAQ label in osteoarthritis model

An Osteoarthritis, Modeling Technology for Genomics Transcriptomics and Proteomics

Inactive Publication Date: 2018-02-02
南宁科城汇信息科技有限公司
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Problems solved by technology

[0010] The method of using iTRAQ markers in the STR / ort model of osteoarthritis is an indispensable part of this research. The technical scheme of the method of using iTRAQ markers in the STR / ort model of osteoarthritis of the present invention has been retrieved There is no similarity in the same industry in China

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Embodiment

[0031] The use of this iTRAQ marker in the STR / ort model of osteoarthritis,

[0032] Consists of the following steps:

[0033] ①Sample preparation:

[0034] 10 STR / ort model mice and 10 control mice were raised until 20 weeks old (adult), and 0.9-1.5 mL of blood was collected from each mouse through the eyeball; after the blood sample was left to stand at 4°C for 1 hour, it was centrifuged at 5000 r / min for 5 minutes , take the supernatant, put 50 μL / tube into a 0.2 mL EP tube, mark it well, and store it at -80°C for later use;

[0035] ② High-performance liquid phase (LC) to remove high-abundance proteins:

[0036] For sample pretreatment, 10 samples of sera from each of the STR / ort model mice and control mice were mixed in equal volumes to form a pool sample of the STR / ort model group and the control group. , bufferA75 μL), a total of 100 μL; after filtration with a 0.22 μm filter membrane, the mixed serum samples of the STR / ort model and the control group were separated ...

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Abstract

The invention discloses a using method of an iTRAQ label in an osteoarthritis model. The using method comprises the steps of 1, preparing samples, 2, conducting efficient liquid chromatography (LC) toremove high-richness protein; 3, adjusting the concentration of a protein sample; 4, labeling an iTRAQ reagent; 5, separating strong cation exchange columns (SCX); 6, conducting point target separation on nano LC, wherein SCX separation components are collected and are afterwards subjected to freeze-drying, and are subjected to separation and point target through reversion phase chromatography; 7, conducting mass spectrometric detection, wherein mass spectrometry is conducted to collect 900-4,000 m / z of atlases, and each target spot is collected 1,500 times in total. By confirming markers ofcrowd samples, OA candidate markers are evaluated, the value of OA is evaluated, the relevancy of the OA and the clinic is analyzed, and the feasibility of the markers in the aspect of clinical application like prevention and early diagnosis of OA and the significance of the markers in crowd bone joint health early-warning are put forward.

Description

technical field [0001] The invention relates to the fields of gene transcriptomics and proteomics, in particular to a method for using iTRAQ markers in osteoarthritis models. Background technique [0002] Osteoarthritis (OA), also known as degenerative bone joint disease, is one of the most common joint diseases, mainly manifested as slowly developing pain, stiffness, swelling of the joints, accompanied by joint dysfunction, and even disability[1- 2]. The pathogenesis of OA is that joints are affected by genetic, biological and mechanical events (such as genetics, metabolic abnormalities, strain, trauma, etc.), which induce changes in cells and matrix in terms of morphology, biomechanics, and molecules. It leads to an imbalance in the degradation and synthesis of articular chondrocytes, extracellular matrix and subchondral bone. The outcome is softening, fibrosis, ulceration and loss of chondrocytes. After losing the protective effect of cartilage, the subchondral bone har...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 于思创王海云
Owner 南宁科城汇信息科技有限公司
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