Protein markers of myocardial infarction in urine and their use in diagnosis and prognosis
A myocardial infarction and application technology, applied in the field of protein markers of myocardial infarction in urine, can solve the problems of inability to detect the disease process and treatment effect in time
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Embodiment 1
[0056] Example 1: Detection of myocardial infarction-related proteins in urine
[0057] We used isotope labeling relative and absolute quantification (iTRAQ) labeling differential proteomics to screen for myocardial infarction-related proteins in urine.
[0058] Materials and Reagents
[0059] 1) Instrument: Triple TOF 5600 mass spectrometer (Absciex Company).
[0060] 2) Main reagents: trypsin (Promega); C18 solid-phase extraction column (3CC, 60mg, Waters); 4 standard iTRAQ reagents (Absciex); C18 reversed-phase chromatography column (4.6mm×250mm, C18, 3μm , Waters Corporation).
[0061] 3) Samples: urine from 7 patients with myocardial infarction (within 12 hours of onset and 7 days after treatment) and 7 normal controls, from the Fourth Affiliated Hospital of Jilin University.
[0062] 1.1 Preparation of urine protein
[0063] Urine samples were centrifuged at 3,500 g for 15 min at 4°C to remove cell debris. Take the supernatant, add pre-cooled absolute ethanol (-20°C...
Embodiment 2
[0070] Example 2: Mass Spectrometric Detection of Urinary Protein Antithrombin-III, Complement C3, α-1-Acid Glycoprotein 1, Serum Transferrin and Cathepsin Z
[0071] In order to better carry out the application detection of these differential proteomic analysis results in further large-scale clinical samples, we used targeted mass spectrometry to monitor the differential proteins identified. Multiple Reaction Monitor (MRM) is a target mass spectrometry quantitative analysis technology based on MS / MS signals, which can simultaneously perform absolute and relative quantitative determinations without the assistance of special chemical labels.
[0072] Materials and Reagents
[0073] 1) Instrument: QTRAP 6500 mass spectrometer (AB Sciex Company).
[0074] 2) Main reagents: trypsin (Promega Company); C18 solid-phase extraction cartridge (3CC, 60 mg, Waters Company).
[0075] 3) Samples: urine from 15 patients with myocardial infarction (within 12 hours of onset and seven days af...
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