MassIVEapplication/xmlftp://massive-ftp.ucsd.edu/v03/MSV000087860/primaryOK2000000ProteomicsChao-Jung ChenamaXis IIautoflex III TOF/TOF smartbeamOrbitrap Exploris 480Homo Sapiens (ncbitaxon:9606)https://massive.ucsd.edu/ProteoSAFe/dataset.jsp?task=dc7acb8622a2431585b1a7df62c64a44China Medical Universitycjchen@mail.cmu.edu.twMassIVE1,283UNIMOD:7 - "Deamidation."MOD:01214 - "modification from UniMod Chemical derivative - OBSOLETE because redundant, the difference component of MOD:01060. Remap to MOD:01060."UNIMOD:35 - "Oxidation or Hydroxylation."The complexity of the proteome often limits the number of identified proteins in the nanoflow LC-MS (nanoLC-MS) analysis of samples. Therefore, peptide fractionation is essential for reducing the sample complexity and improving the proteome coverage. In this study, to achieve high-pH reversed-phase (RP)-well plate fractionation for high-throughput proteomics analysis, C18 particles were coated on a 96-well plate, and the sample-loading processes were optimized for high-pH fractionation. The sample capacity of the high-pH RP-well plate was estimated to be ~6 μg of protein. There were 1.85- and 1.71-fold increases in the number of protein groups and peptides identified, respectively, with high-pH RP-well plate fractionation, compared to those without fractionation. In addition, with alkaline C18 well plate fractionation, exosome markers could be detected using ~1 μg of a protein digest of exosomes by microflow LC-MS (microLC-MS). These results illustrate that high-pH RP-well plate fractionation has superior sensitivity and effectiveness in preparing trace amounts of proteins for deep proteome analysis.Development of a high-pH reversed-phase well plate for peptide fractionation and deep proteome analysis of cells and exosomes.Liu Yu-Ching YC, Huang Yu-Ting YT, Chen Chao-Jung CJdevelopment, polypeptide, peptide, Polypeptides, peptido, peptides, determination, chemical analysis, postnatal development, postnatal growth, Peptid, peptidos, total expressed protein, Cell., Polypeptide, assay, growth and development, backward, growth, Proteomes, Peptide, reversedprojections, determination, Peptidomics, lamellae, protein complex, Ass-1, Proteins, Liquid Chromatography Mass Spectrometry, lamina, total expressed protein, High Performance Liquid Chromatography Mass Spectrometry, flanges, Gene, Ultra Performance Liquid Chromatography-Mass Spectrometry, High Pressure Liquid Chromatography-Mass Spectrometry, protein, protein-containing complex, process of organ, Peptide, High Performance Liquid Chromatography-Mass Spectrometry, polypeptide, peptide, lamella, Polypeptides, protein polypeptide chains, HPLC-MS, Liquid Chromatography-Mass, peptido, native protein, natural protein, polypeptide chain, C18, sensitive, AA408052, Protein, chemical analysis, B1, shelf, Gene Products, fold, extracellular vesicular exosome, protein aggregate, sensitivity, flange, organ process, UPLC-MS, study, Ultra Performance Liquid Chromatography Mass Spectrometry, Ultra-Performance Liquid Chromatography-Mass Spectrometry, peptides, High Pressure Liquid Chromatography Mass Spectrometry, shelves, Spectrometry, Specificity, chemical analysis., Spectrometries, proteins, ridges, projection, ridge, sample population, Protein Gene Products, ASS, allergic reaction, Gene Proteins, processes, Specificity and Sensitivity, Liquid Chromatography-Mass Spectrometries, papilla, sample, D1, Chromatography-Mass Spectrometry, Liquid, LC-MS, Sensitivity, Peptid, peptidos, assay, Polypeptide, Chromatography-Mass Spectrometries, laminae, Proteomes, PTHB1, exosomedevelopment, polypeptide, peptide, Polypeptides, peptido, peptides, determination, chemical analysis, postnatal development, postnatal growth, Peptid, peptidos, total expressed protein, Cell., Polypeptide, assay, growth and development, growth, Proteomes, Peptidedevelopment, polypeptide, peptide, Polypeptides, peptido, peptides, determination, chemical analysis, postnatal development, postnatal growth, Peptid, peptidos, total expressed protein, Cell., Polypeptide, assay, growth and development, backward, growth, Proteomes, Peptide, reversed0falseDevelopment of a high-pH reversed-phase well plate for peptide fractionation and deep proteome analysis of cells and exosomesDevelopment of a high-pH reversed-phase well plate for peptide fractionation and deep proteome analysis of cells and exosomesWed Jul 21 02:45:00 BST 2021MSV00008786035099582