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Protein amino acid sequence de novo sequencing method based on unequal stable isotope labeling at two ends of polypeptide

ActiveCN105301119AHigh reaction efficiency and selectivityHigh specificityComponent separationChemistrySide chain
The invention relates to a protein amino acid sequence de novo sequencing method based on unequal stable isotope labeling at two ends of polypeptide. The method comprises the following steps: conducting enzyme digestion on protein subjected to denatured reduced alkylation by intracellular protease lysine-C to form peptide fragments containing N-terminal alpha-amino and C-terminal lysine side chain epsilon-amino; after the sample is divided into two parts, conducting selective derivatization on the N-terminal alpha-amino of the polypeptide by using a labeling reagent containing different isotopes; then conducting derivatization on C-terminal lysine side chain epsilon-amino of the polypeptide by using the labeling reagent containing different isotopes, or in cell culture, labeling lysine in the protein by using a culture solution containing isotope labeling lysine; finally mixing the sample, and conducting separation and identification by using high performance liquid chromatography- mass spectrometry. According to the method, paired polypeptide fragment ions are formed through labeling for resolution, the influence of interference signals is reduced, the selective specificity of the fragment ions and the accuracy of polypeptide sequencing are improved, and the speed of de novo sequencing is improved.
Owner:DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI

Ratio-type nano silicon quantum dot fluorescence probe and preparation method and application thereof

The invention belongs to the technical field of nano material science and molecular biology, and particularly relates to a ratio-type nano silicon quantum dot fluorescence probe and a preparation method and application thereof. The preparation method comprises the steps that a one-pot method is adopted, triaminopropyl-triethoxysilane is reduced with sodium ascorbate at room temperature, and water-soluble silicon quantum dots are obtained; the water-soluble silicon quantum dots and chlorin e6 (Ce6) are mixed and subjected to standing at the room temperature under the lucifugal condition, and a silicon quantum dot and chlorin e6 composite material is obtained. The composite material emits fluorescence with the wavelength of 430 nm-580 nm and the wavelength of 640 nm-680 nm under the fluorescence excitation of 410 nm, and active oxygen radicals can specifically quench fluorescence of silicon quanta and basically have no influence on fluorescence of Ce6. Therefore, the composite material can serve as a ratio-type nano silicon quantum dot fluorescence probe to be used for detecting the content of hydroxyl radicals (-OH), rapidness and high efficiency are achieved, the specificity is high, the sensitivity is high, the probe is used for in-vivo cell imaging, the biocompatibility is good, and the visual degree is high.
Owner:FUDAN UNIV
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