Istically significant was defined as p <0.05.Western blot analysis for validationWe > 자유게시판

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Istically significant was defined as p < 0.05.Western blot analysis for validationWe utilized a nano-liquid chromatography lectro spray ionization andem mass spectrometry (nano-LC/ESI?mass spectrometry/mass spectrometry) system consisting of PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/14960617 an Aquity UPLC system (Waters Corporation, Milford, USA) and a LTQ Obitrap XL mass spectrometer (Thermo Fisher, waltham, MA,USA) equipped with a nano-ESI source(Michrom Bioresources, Auburn, USA) to perform peptide sequencing and peptide identification. The peptide solutions purified by magnetic beads, were loaded to a C18 trap column (nanoACQUITY) (180 m ?20 mm ?5 m (symmetry)). The flow rate was 15 l/min. Then the desalted peptides were analyzed by C18 analytical column (nano ACQUITY) (75 m ?150 mm ?3.5 m (symmetry)) at a flow rate of 400 nl/min for 60 min. TheSodium dodecyl PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/21715270 sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting were performed essentially as described elsewhere. Briefly, cell pellets were resuspended on ice in lysis buffer containing 10 mM Tris Cl (pH 7.4), 5 mM MgCl2, 1 Triton X-100, 100 mM NaCl, 10 mM NaF, 1 mM Na3VO4 and a protease inhibitor cocktail. After sonication, cellular proteins were separated on an SDS-polyacrylamide gel and transferred to polyvinylidene fluoride membranes (Roche Diagnostics Corporation, Indianapolis, Indiana United States), which were probed with the appropriate primary antibodies. Immunoreactivity was detected with the relevant horseradish peroxidase-labeled secondary anti-bodies which, in turn, were visualized on an Image Reader Tano-5500 (Tano, Shanghai, Methyl 5-amino-2,4-difluorobenzoate China) using chemiluminescence substrate reagent purchased from 7 sea pharmtech (Shanghai, China) For quantification of the data, the images were further analyzed on the same instrument using 2D Densitometry Image Analyzer IPP 7.0 software (Tano, Shanghai, China).Bai et al. Proteome Science 2013, 11:39 http://www.proteomesci.com/content/11/1/Page 15 ofAdditional fileAdditional file 1: Figure S1. Serum peptide fingerprints of the same serum through same processing in three-repeated experiments. (A) Serum peptide fingerprints of the same acute leukemia through same processing in three-repeated experiments. (B) Serum peptide fingerprints of the same healthy control through same processing in three-repeated experiments.8.9.10.Abbreviations MRD: Minimal residual disease; MB-WCX: Weak cation exchange magnetic beads; MALDI-TOF-MS: Matrix assisted laser desorption ionization time of flight mass spectrometry; AML: Acute myeloid leukemia; HPLC-MS/MS: High-performance liquid chromatography tandem mass spectrometry/mass spectrometry; CR: Complete remission; QC: 3-(tert-Butyldimethylsilyloxy)propan-1-amine Quick classifier; UBA1: Ubiquitinlike modifier activating enzyme 1; PF4: Platelet factor 4; RTQ-PCR: Real-time quantitative polymerase chain reaction; SELDI-TOF-MS: Surface enhanced laser desorption ionization time of flight mass spectrometry; 2-DE: Two-dimensional gel electrophoresis, Rho-GDP; MM: Multiple myeloma; SNN: Supervised neural network; CV: Coefficient of variation; MW: Molecular weight; GA: Genetic algorithm; ELISA: Enzyme linked immunosorbent assay; OS: Overall survival; HPV: Human papilloma virus. Competing interests The authors declare no conflict of interests with any company or financial organization. Authors' contributions JB and YZ were involved in serum peptide purification, writing the draft of the manuscript; JY performed serum peptide profiling data acquisition and processing; YY and JLW were involved in serum s.