Mass spectra identification method of nucleophosmin variable spliceosome and gastric cancer diagnostic reagent kit

The technology of a nucleolar phosphoprotein and an identification method is applied in the detection field of biological samples, which can solve the problems of poor repeatability and high randomness of experiments, and achieve the effect of good repeatability and reliable identification results.

Active Publication Date: 2011-08-17
BEIJING 3S CENTURY TECH CORP
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Problems solved by technology

Although Maldi-TOF is widely used, in practice, due to the limitation of its principle, it has great selectivity to the excited fragments, and many fragments will not appear in the mass

Method used

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  • Mass spectra identification method of nucleophosmin variable spliceosome and gastric cancer diagnostic reagent kit
  • Mass spectra identification method of nucleophosmin variable spliceosome and gastric cancer diagnostic reagent kit
  • Mass spectra identification method of nucleophosmin variable spliceosome and gastric cancer diagnostic reagent kit

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Embodiment

[0052] Specimen collection and processing:

[0053] Within 10 minutes after surgical resection, the specimens were immediately placed in liquid nitrogen and stored at -80°C until protein extraction. The full-thickness gastric tissue (more than 6 cm from the tumor edge) not invaded by the tumor was taken from the same patient as a normal control, and different instruments were used during the collection process to prevent cross-contamination of the specimens.

[0054] 1. Protein extraction

[0055] Protein was extracted using Trizol kit according to the instructions. Specific steps are as follows:

[0056] (1) Add 1ml Trizol reagent to 50mg frozen tissue for lysis, shake vigorously to lyse the cells;

[0057] (2) Transfer the above-mentioned Trizol lysate into a centrifuge tube, and place it at a room temperature of 15°C to 30°C for 5 minutes;

[0058] (3) Add 0.2ml chloroform to each 1ml Trizol lysate in the above centrifuge tube, cover the tube cap, shake vigorously for 1...

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Abstract

The invention provides a mass spectrometry identification method of nucleophosmin variable spliceosome and a gastric cancer diagnostic reagent kit. The method is combined by a Glu-C enzyme cutting method and a Q-TOF (quadrupole time-of flight) mass spectra method, and can be applied to the clinical diagnosis and treatment evaluation of the gastric cancer; and by the method disclosed by the invention, the rapid and reliable identification result can be acquired as compared with the traditional methods.

Description

technical field [0001] The invention relates to the detection field of biological samples, in particular to a mass spectrometry identification method for mutant proteins. Background technique [0002] Nucleophosmin (nucleophosmin, NPM, B23, N038 or NPM1) is one of the main protein molecules located in the nucleolar granule region, the human B23 gene is located on chromosome 5q35, and the gene is about 23kb in length (Chan, P.K., 1986, Zhang, X.X , 1989; Cochrane, A.W, 1990), including 12 exons, encoding a protein consisting of 294 amino acids. Under normal circumstances, nucleolin phosphoprotein is mainly located in the nucleolar granule region, but it can play an important role by shuttling in the nucleolus, nucleoplasm and cytoplasm (Charles G, 2008). [0003] The sequences of the 12 exons (Exon1-12) of nucleolin are as follows: [0004] [0005] [0006] The currently recognized B23 gene has two transcriptional splicing forms. The B23 protein was first reported b...

Claims

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Application Information

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IPC IPC(8): G01N27/62G01N30/02G01N30/06C07K7/06
Inventor 季加孚彭勇张连海李子禹宗祥龙吴晓江胡颖
Owner BEIJING 3S CENTURY TECH CORP
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