JPOST Repositoryapplication/xmlhttps://storage.jpostdb.org/JPST000315/files/parameters.txthttps://storage.jpostdb.org/JPST000315/files/peptides.txthttps://storage.jpostdb.org/JPST000315/files/summary.txthttps://storage.jpostdb.org/JPST000315/files/proteinGroups.txthttps://storage.jpostdb.org/JPST000315/files/3Flag_only_CLIP_3.rawhttps://storage.jpostdb.org/JPST000315/files/3Flag_UBL3_CLIP_7.rawhttps://storage.jpostdb.org/JPST000315/files/3Flag_UBL3C113114A_CLIP_12.rawhttps://storage.jpostdb.org/JPST000315/files/3Flag_only_CLIP_4.rawhttps://storage.jpostdb.org/JPST000315/files/3Flag_only_CLIP_2.rawhttps://storage.jpostdb.org/JPST000315/files/3Flag_UBL3_CLIP_8.rawhttps://storage.jpostdb.org/JPST000315/files/3Flag_UBL3C113114A_CLIP_11.rawhttps://storage.jpostdb.org/JPST000315/files/3Flag_UBL3C113114A_CLIP_13.rawhttps://storage.jpostdb.org/JPST000315/files/3Flag_UBL3_CLIP_6.rawprimaryOK200ProteomicsKunihiro TsuchidaHomo Sapiens (human)https://repository.jpostdb.org/entry/JPST000315Fujita Health UniversityjPOSTDmelCG9038, HCG-1, CG9038, hcg-1, AW108023, MUB, Ubl3, pnsc1, HCG, PNSC1.LF 2000, IL1BC, T22F8.160, MeCN, Meth, NCMe, and rna binding 2, Laundry Pods, Chronic Airflow Obstructions, Sepharose C1 4B, Synthetic Detergents, TFA, CASP-1, classic hairy cell leukaemia, CG7788, crice, Syndet, chronic obstructive pulmonary disease (COPD), protein, Hydrogenchlorid, Basodexan, Trifluoroacetate, Detergent, Polypeptides, Pod, Synthetic Detergent, Triton, hydrogen chloride, peptido, T22F8_160, Detergents, ATGPR7, CH3-C#N, chronic obstructive airways disease NOS (disorder), Chronic irreversible airway obstruction, SEC, DGS, GRP8, GRP7, Min, Analysis, Chronic Obstructive Pulmonary Disease (COPD), protein aggregate, Il1bc, Nereid, DmelCG42273, Mass Spectrum Analysis, average, Sepharose, COLD (chronic obstructive lung disease), peptides, Analyses, NEC in ICD9CM_2006, DmelCG13176, E927b, sec, min, CAFL - Chronic airflow limitation, ORF19, mAPC, IBP, IL-1BC, Chronic airflow limitation, AI047805, Wasserstoffchlorid, C1 4B, ICE, Ice, disease (COPD), GLYCINE RICH PROTEIN 7, DmelCG1106, trifluoro-, TF, autolysin activity, necrosis, Bladder pain syndrome, glycine-rich RNA-binding protein 8, CHRONIC OBSTRUCTIVE PULMONARY DISEASE, CHRONIC, Detergent Pod, chronic obstructive lung disease [Ambiguous], obstructive lung disease, anatomical protrusion, Caspase-1 subunit p10, BZRP, iCE, ice, drice, 4B, Dmel_CG7826, Normalcy, classic hairy cell leukemia, LF2000, Spectrum Analysis, Airflow Obstructions, ur, DmelCG7788, PBS, PBR, Spectroscopy, CHRONIC OBSTRUCTIVE AIRWAY DIS, pulmonary disease (COPD), carbamide, Dm1, drIce, drICE, [4)-3, CHRONIC OBSTRUCTIVE, chronic obstructive pulmonary disease and allied conditions, Sepharose C1, Chlorwasserstoff, Chronic airway disease, bacteriocin activity, Laundry Detergent Pod, NOS, Cleansing Agents, 1110049F14Rik, Individual, WASH, Airflow Obstruction, Detergent Pods, Dmel_CG7835, Acetic acid, COPD NOS, Crystal, CHRONIC OBSTRUCTIVE LUNG DIS, GLYCINE-RICH PROTEIN 8, Mnb, MNB, Chronic Obstructive Airways Disease, Chronic airway obstruction, Acid, Chronic obstructive lung disease, absence of, Painful Bladder Syndrome, Pods, cold, P45, HCl, Trifluoroacetic, Spectrometry, AW124434, beta Trypsin, CG1106, Speed, Chronic Airflow Obstruction, H2NC(O)NH2, Health, lysin activity, Painful bladder syndrome, F2G1.4, Agents, Cesium, OBSTRUCTIVE PULMONARY DISEASE (COPD), spine, COPD - Chronic obstructive pulmonary disease, Acetamide, Drice, chronic obstructive airways disease, Sepharose 4B, p45, bacteriolytic toxin activity, Polypeptide, obstructive pulmonary disease (COPD), methyl cyanide, chlorane, DRICE, chronic obstructive lung disease, COLD, 1728, Bladder Pain Syndrome, BPBS, Laundry Detergent Pods, chlorure d'hydrogene, DISEASE (COPD), DrIce, DrICE, CAO - Chronic airflow obstruction, Gene, Carbamide, circadian rhythm, CHRONIC OBSTRUCTIVE PULM DIS, Spectrum Analyses, Chronic obstructive pulmonary disease NOS, abdominal muscles, protein-containing complex, Chalk, Normalcies, and RNA binding 1, CG7826, Buffer, protrusion, Chronic obstructive pulmonary disease finding, Agent, Karbamid, Cold, PULMONARY DISEASE (COPD), Pulmonary Disease, p63, p65, Harnstoff, Tina, CG7835, Gene Products, Mass, CG42273, Carmol, cloruro de hidrogeno, Mass Spectroscopy, Pro-Mega, Normalities, Cesium Trifluoroacetate, T6K12_14, Synthetic, Cold Temperatures, Glass, FAM39E, Hydrogen chloride, gel, Selb, PCE-2, beta-Trypsin, pk18, Chronic Obstructive, MBR, Laundry Detergent, Caspase-1 subunit p20, ATGRP7, ATGRP8, NEC, Agarose, LF-2000, PTBR, Chronic, Laundry Pod, peptidos, 6-An-alpha-L-Galp-(1->3)-beta-D-Galp-(1->]n, 2-iodo-, MDA-MB-231, Laundry, COPD, Individual Health, COAD - Chronic obstructive airways disease, DYRK1, 1-(14)C-labeled, chronic obstructive airways disease NOS, DBI, 3H-labeled, caspase 3, protein complex, not elsewhere classified, COAD, Proteins, chloridohydrogen, CE-2, COLD - Chronic obstructive lung disease, acetonitrile, buffer, Cell, chronic, PULM DIS CHRONIC OBSTRUCTIVE, Peptide, Chronic obstructive pulmonary disease finding (finding), CD142, [HCl], Wash2, Wash1, secret agent, MS, Temperatures, native protein, PKBS, carbonyldiamide, Protein, Hydrochloride, Dyrk1, CES2A1, GLYCINE-RICH RNA-BINDING PROTEIN 7, (COPD), chronic airway obstruction, eagle-Barrett syndrome, chronic obstructive, ACETONITRILE, CAL - Chronic airflow limitation, Mass Spectrum Analyses, cyanomethane, CG13176, Cleansing, T6K12.14, chronic obstructive airway disease, AU020952, Temperature, Mass Spectrum, with urinary tract Abnormality and cryptorchidism, holin, PRSS, Normality, uree, Dops, CC1, GR-RBP7, GR-RBP8, Tripcellim, lipofectamine 2000, IL-1 beta-converting enzyme, ethanenitrile, Episome, Interleukin-1 beta-converting enzyme, TFA., mDRC, ME-IV, Interleukin-1 beta convertase, Protein Gene Products, Episomes, Gene Proteins, Trypure, Cleansing Agent, chronic obstructive lung disease (disorder), 3.4.22.36, Plasmid, cold (chronic obstructive lung disease), Peptid, Chronic Obstructive Lung Disease, hydrochloric acid, chronic obstructive pulmonary diseasefalseFLAG-IP-UBL3MDA-MB-231 cells were transfected with plasmids using Lipofectamine 2000. After 24 hr, cells were washed and collected by scraping into ice-cold PBS, pelleted by centrifugation at 400 × g for 1 min at 4°C, and lysed with 1% Triton buffer for 20 min at 4°C. Crude nuclei and unbroken cells were excluded by centrifugation at 20,000 × g for 5 min at 4°C. As a preclean process, the supernatants were incubated twice with 40 μL Protein G-sepharose beads (GE Health care, 50% slurry in lysis buffer) for 1 hr at 4°C. The supernatants were incubated with 50 μl anti-FLAG M2 affinity gel (Sigma, 50% slurry in lysis buffer) for 16hr at 4°C. The beads were washed with 1% Triton buffer and then three times with wash buffer (50mM Tris-HCl [pH7.5]) to remove detergent, and resuspended in 200 μL digestion buffer (2M urea, 50mM Tris-HCl [pH 7.4], 1mM DTT, 5mM iodoacetamide, 1 μg of trypsin [Promega, V5280]), and incubated by shaking at 1,200 rpm for 16hr at 37°C. The samples were acidified with 22 μL of 10% trifluoroacetic acid (TFA). The supernatant containing peptides were then taken by centrifugation at 4,400 × g for 30 sec. The peptides were desalted by C18Tip (Thermo, StageTips), dried by speedvac, and then resuspended in mass spectrometry buffer (2% acetonitrile, 0.2% TFA).Mon Sep 03 00:00:00 BST 2018PXD0076179606