Application of urine protein marker in preparation of kit for detecting clinical stages of diabetic nephropathy
A technique for diabetic nephropathy and clinical staging, which is applied in the field of diabetic nephropathy and can solve problems such as deviation
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[0061] Part I: Urine Protein Sample Preparation
[0062] Example 1 of an invention patent application whose publication number is CN 108333263 A and whose publication date is July 27, 2018. The preparation method is prepared.
[0063] Part II: Mass Spectrometry Detection
[0064] Digested samples were separated from tryptic peptides on a homemade capillary column packed with C18 particles and analyzed by a Thermo Fisher Orbitrap mass spectrometer coupled to an online Easy-nLC 1000 Nano-HPLC system (Thermo Fisher Scientific). LC-MS / MS data were processed in the FIRMIANA analysis platform [Nat Biotechnol.2017 May 9;35(5):409-412.], and protein searches were performed on the Mascot search engine.
[0065] Protein abundance was measured as iBAQ (Intensity-Based Relative Quantification)-label-free quantification algorithm. For batch-to-batch comparisons, iBAQ was converted to iFOT (intensity-based total fraction), representing the normalized intensity of proteins identified in t...
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