Comprehensive and Reproducible Phosphopeptide Enrichment Using Iron Immobilized Metal Ion Affinity Chromatography (Fe-IMAC) Columns

Name: Work Video:
Duration: 3 min 30 s
Benjamin Ruprecht; Heiner Koch; Guillaume Médard; Max Mundt; Bernhard Küster; Simone Lemeer
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1100, 2899, 2912], 'enable': [5, 277], 'the': [6, 53, 70, 138, 179, 197, 205, 210, 228, 238, 241, 278, 325, 342, 410, 451, 469, 477, 482, 500, 510, 513, 683, 712, 816, 943, 994, 1003, 1025, 1108, 1132, 1177, 1260, 1293, 1318, 1346, 1362, 1366, 1410, 1627, 1631, 1642, 1670, 1824, 1832, 1852, 1857, 1886, 1911, 2089, 2371, 2441, 2452, 2557, 2763, 2798, 2906, 2930, 3031, 3068, 3135, 3169, 3306, 3362, 3379, 3477, 3481, 3516, 3532, 3573, 3636, 3658, 3743, 3800, 3854, 3880, 3885, 4043, 4058, 4067, 4081, 4130], 'identification': [7, 166, 279, 438, 1026, 1133, 1261, 2323, 2372], 'of': [8, 10, 55, 76, 79, 100, 123, 127, 164, 171, 184, 209, 231, 240, 269, 280, 282, 327, 348, 351, 372, 395, 399, 436, 443, 456, 481, 503, 512, 541, 595, 611, 682, 696, 698, 707, 716, 797, 818, 836, 862, 903, 946, 975, 1002, 1027, 1073, 1103, 1107, 1134, 1172, 1176, 1262, 1297, 1321, 1365, 1432, 1467, 1499, 1531, 1562, 1593, 1633, 1665, 1669, 1692, 1720, 1743, 1785, 1808, 1826, 1834, 1851, 1888, 1894, 1914, 2084, 2223, 2241, 2283, 2324, 2373, 2445, 2451, 2469, 2500, 2539, 2562, 2582, 2613, 2732, 2864, 2886, 2909, 2918, 2934, 3084, 3115, 3185, 3208, 3260, 3275, 3302, 3333, 3347, 3355, 3364, 3367, 3387, 3400, 3422, 3489, 3500, 3522, 3544, 3552, 3559, 3677, 3750, 3755, 3835, 3846, 3853, 3873, 3884, 3893, 3909, 3927, 3934, 3974, 4012, 4025, 4053, 4060, 4063, 4090, 4099, 4114, 4132, 4135], 'thousands': [9, 281, 697], 'phosphopeptides': [11, 77, 113, 129, 283, 349, 385, 401, 904, 947, 1028, 1135, 1239, 1263, 1915, 2325, 2374, 2446, 2865], 'from': [12, 42, 87, 94, 284, 314, 359, 366, 948, 1029, 1136, 1264, 1312, 1323, 1401, 1435, 1874, 1916, 2219, 2375, 2735, 3531, 3609], 'complex': [13, 80, 285, 352, 1468, 1500, 1532, 2470, 2583, 3975], 'samples.': [14, 286], 'Current': [15, 287], 'offline': [16, 288, 2861], 'approaches': [18, 290, 828], 'using': [19, 249, 291, 521, 951, 1034, 1141, 1269, 1438, 1919, 1986, 2380, 2717, 2741, 2866, 3030, 3212, 3397, 3496, 3710, 3782], 'TiO2,': [20, 292, 2540], 'Ti,': [21, 293], 'and': [22, 44, 73, 91, 117, 155, 167, 190, 200, 227, 257, 294, 316, 345, 363, 389, 427, 439, 462, 472, 499, 529, 565, 694, 711, 873, 905, 953, 991, 993, 1065, 1075, 1356, 1462, 1474, 1494, 1506, 1526, 1538, 1579, 1640, 1651, 1806, 1870, 1921, 2015, 2131, 2247, 2286, 2322, 2349, 2425, 2434, 2464, 2476, 2531, 2542, 2568, 2577, 2589, 2656, 2692, 2766, 2856, 2877, 2880, 2897, 2901, 2923, 2954, 2987, 3006, 3055, 3100, 3172, 3177, 3206, 3283, 3308, 3338, 3352, 3390, 3409, 3431, 3494, 3527, 3547, 3556, 3570, 3585, 3631, 3650, 3665, 3731, 3746, 3830, 3857, 3876, 3906, 3931, 3942, 3948, 3962, 3969, 3981, 4019, 4042, 4056, 4093, 4140], 'Fe': [23, 295], 'immobilized': [24, 296, 823, 839, 919, 1214, 1949, 2049, 2159, 2183, 3713, 3785], 'metal': [25, 297, 738, 758, 840, 920, 925, 1067, 1215, 1950, 2050, 2160, 2184], 'ion': [26, 298, 739, 759, 921, 1068, 1216, 2161, 2185, 3716, 3788], 'affinity': [27, 299, 740, 760, 817, 842, 860, 922, 927, 1069, 1217, 1951, 2051, 2162, 2186, 3265, 3717, 3789], 'chromatography': [28, 300, 861, 923, 928, 1070, 2052, 2163, 2173, 2187, 2197, 2230, 2245, 2252, 2271, 2336], '(IMAC)': [29, 301], 'material': [30, 302, 3381, 3856], 'batch': [32, 158, 304, 430, 1384, 3224, 3954], 'or': [33, 159, 305, 431, 1386, 1892, 3442], 'microtip': [34, 306], 'format': [35, 252, 307, 524, 1385, 2919], 'are': [36, 237, 308, 509, 599, 702, 1101, 1379, 2128], 'widely': [37, 309], 'used,': [38, 310, 2427], 'but': [39, 311, 2797], 'they': [40, 312, 1872, 1880], 'suffer': [41, 86, 313, 358, 1400, 1873], 'irreproducibility': [43, 315], 'compromised': [45, 317], 'selectivity.': [46, 318], 'To': [47, 319, 3508], 'address': [48, 320, 2203], 'these': [49, 321, 1098, 1847, 1864], 'shortcomings,': [50, 322], 'we': [51, 323, 1371, 2852, 3514], 'revisited': [52, 324], 'merits': [54, 326], 'performing': [56, 328], 'an': [60, 105, 250, 332, 377, 522, 2210, 3394, 3434, 3454, 3625], 'HPLC': [61, 251, 333, 523, 3455, 3533, 3536], 'column': [62, 334, 3437, 3470, 3482, 3487, 3498, 3517, 3520, 3568, 3637, 3671], 'format.': [63, 335], 'We': [64, 336, 2889, 3087], 'found': [65, 337, 1398, 1620, 3741], 'that': [66, 174, 222, 261, 338, 446, 494, 533, 1097, 1349, 1370, 1618, 2556, 2757, 2891, 2905, 3375], 'Fe-IMAC': [67, 151, 246, 339, 423, 518, 1289, 2870, 3223, 3261, 3327, 3348, 3423, 3436, 3592, 3613, 3641], 'columns': [68, 152, 340, 424, 3139, 3887], 'enabled': [69, 341], 'selective,': [71, 343, 2875], 'comprehensive,': [72, 344, 2876], 'reproducible': [74, 346, 1463, 1495, 1527, 2465, 2578, 2878, 3970], 'out': [78, 350, 2756, 3510, 3590], 'lysates.': [81, 353], 'Column': [82, 354], 'did': [84, 193, 356, 465], 'not': [85, 357, 1094], 'bead-to-sample': [88, 360], 'ratio': [89, 361, 1632, 1644, 1723], 'issues': [90, 362], 'scaled': [92, 364], 'linearly': [93, 365], '100': [95, 367, 3188, 3258, 3504], 'μg': [96, 368, 3189, 3754], 'to': [97, 133, 143, 213, 265, 369, 405, 415, 485, 537, 572, 593, 601, 704, 721, 1233, 1399, 1472, 1504, 1536, 1612, 1621, 1636, 1845, 2021, 2202, 2474, 2587, 2770, 2802, 2975, 3079, 3103, 3130, 3133, 3168, 3187, 3453, 3476, 3562, 3611, 3627, 3657, 3752, 3826, 3979], '5': [98, 370, 2972, 3860], 'mg': [99, 371, 3845], 'digest.': [101, 373], 'Direct': [102, 374], 'measurements': [103, 375, 2778], 'on': [104, 376, 824, 1626, 1724, 1814, 2860, 3331, 3378, 3851], 'Orbitrap': [106, 378], 'Velos': [107, 379], 'mass': [108, 235, 380, 507, 726, 734, 754, 979, 1955, 2082, 2154, 2178, 2337, 2412, 2759, 3246], 'spectrometer': [109, 381], 'identified': [110, 382], '>7500': [111, 383], 'unique': [112, 128, 384, 400, 1111], 'with': [114, 386, 551, 1110, 1292, 1747, 1856, 2333, 2948, 2967, 2984, 3045, 3057, 3075, 3128, 3272, 3323, 3384, 3464, 3473, 3485, 3518, 3541, 3549, 3591, 3605, 3640, 3728, 3832, 3859, 3890, 3924, 3994, 4009, 4022, 4077, 4087, 4096, 4111], '90%': [115, 387], 'selectivity': [116, 388, 1652], 'good': [118, 390], 'quantitative': [119, 391, 1569, 1600, 1699, 2507, 2620], 'reproducibility': [120, 392], '(median': [121, 393], 'cv': [122, 394], '15%).': [124, 396], 'The': [125, 245, 397, 517, 609, 776, 1342, 2403, 2829, 3012, 3025, 3041, 3070, 3237, 3414, 3469, 3535, 3670, 4125], 'number': [126, 398], 'could': [130, 402], 'be': [131, 214, 263, 403, 486, 535, 602, 1234, 2432, 2561, 2636, 3740], 'increased': [132, 404], 'more': [134, 406, 1294, 1838], 'than': [135, 407, 1240, 2786, 2929], '14,000': [136, 408], 'when': [137, 409], 'IMAC': [139, 411, 1038, 1145, 1273, 1745, 2384], 'eluate': [140, 412], 'was': [141, 413, 1619, 3014, 3028, 3043, 3073, 3117, 3290, 3359, 3417, 3471, 3483, 3575, 3578, 3603, 3638, 3672, 3757, 3804, 3956, 4045, 4069, 4127], 'subjected': [142, 414], 'a': [144, 203, 266, 416, 475, 538, 547, 1000, 1314, 1368, 1383, 1563, 1594, 1623, 1647, 1693, 1782, 1786, 1803, 1815, 1843, 2438, 2501, 2537, 2614, 2854, 2867, 2883, 2916, 3080, 3284, 3300, 3465, 3606, 3632, 3651, 3675, 3827], 'subsequent': [145, 417], 'hydrophilic': [146, 418, 2167, 2191, 2227, 2269], 'strong': [147, 419, 743, 763, 2168, 2192, 2248, 2302, 2350], 'anion': [148, 420, 744, 764, 2169, 2193, 2303, 2329], 'exchange': [149, 421, 2170, 2194, 2250, 2304, 2330, 2352], 'separation.': [150, 422], 'outperformed': [153, 425, 2894], 'Ti-IMAC': [154, 426, 1230, 2896, 3680, 3836, 3847], 'TiO2': [156, 428, 1241, 1564, 1595, 1694, 2502, 2615, 2898, 3953, 3997], 'tip': [160, 432], 'mode': [161, 433], 'terms': [163, 435], 'intensity.': [168, 440], 'Permutation': [169, 441], 'enrichments': [170, 442, 1378, 3225], 'flow-throughs': [172, 444], 'showed': [173, 445], 'all': [175, 447, 596, 2895, 2910, 2959], 'materials': [176, 199, 211, 448, 471, 483], 'largely': [177, 449], 'bound': [178, 450], 'same': [180, 452], 'species,': [182, 454], 'independent': [183, 455], 'physicochemical': [185, 457, 1112, 2932], 'characteristics.': [186, 458], 'However,': [187, 459, 2753], 'binding': [188, 460, 3276, 3288, 3297, 3349, 3874, 4026, 4054, 4091], 'capacity': [189, 461], 'elution': [191, 463, 1311, 2571, 3602, 3943, 4115], 'efficiency': [192, 464], 'profoundly': [194, 466], 'differ': [195, 467], 'among': [196, 468], 'formats.': [201, 473], 'As': [202, 474, 1842, 2437], 'result,': [204, 476], 'often': [206, 478, 713, 1091, 1646, 1829, 2208], 'quoted': [207, 479], 'orthogonality': [208, 480, 2908], 'has': [212, 484, 1860, 2455], 'called': [215, 487], 'into': [216, 488, 1391, 3393], 'question.': [217, 489], 'Our': [218, 490], 'results': [219, 491], 'strongly': [220, 492], 'suggest': [221, 493], 'insufficient': [223, 495, 2924], 'capacity,': [224, 496, 1876, 2567], 'inefficient': [225, 497, 2921], 'elution,': [226, 498, 2922], 'stochastic': [229, 501], 'nature': [230, 502], 'data-dependent': [232, 504, 2803], 'acquisition': [233, 505, 2926], 'spectrometry': [236, 508, 727, 1956, 2083, 2155, 2179], 'causes': [239, 511], 'experimentally': [242, 514], 'observed': [243, 515], 'complementarity.': [244, 516], 'workflow': [248, 520, 2858], 'developed': [253, 525], 'here': [254, 526], 'enables': [255, 527], 'rapid': [256, 528, 1319], 'comprehensive': [258, 530, 1363, 1783], 'phosphoproteome': [259, 531, 1109, 2255, 3708, 3780], 'analysis': [260, 532, 722, 796, 1887, 2410, 2696, 3244], 'can': [262, 534, 1610, 2431, 2635, 3739], 'applied': [264, 536, 2020], 'wide': [267, 539, 2884], 'range': [268, 540, 2768, 2885], 'biological': [270, 542, 563], 'systems.': [271, 543], 'Protein': [544, 3036], 'phosphorylation': [545, 568, 700, 798, 1961, 2024, 2086, 2409, 3243], 'is': [546, 558, 569, 769, 999, 1090, 1231, 1290, 1348, 1352, 1359, 1630, 1645, 1811, 1878, 2800, 2913], 'reversible': [548], 'post-translational': [549], 'modification': [550], 'pivotal': [552], 'roles': [553], 'cellular': [555], 'signaling.': [556], 'It': [557, 1089], 'implicated': [559], 'many': [561], 'essential': [562], 'processes,': [564], 'aberrant': [566], 'protein': [567, 612, 1637, 1960, 2733, 3026, 3071], 'causally': [570], 'linked': [571], 'numerous': [573], 'diseases': [574], '(1.Blume-Jensen': [575], 'P.': [576, 608, 857, 1426, 1556, 1587, 1686, 2494, 2520, 2607, 2648, 2726], 'Hunter': [577], 'T.': [578, 657, 1017, 1124, 1252, 2363, 2526, 2644, 2724], 'Oncogenic': [579], 'kinase': [580, 1032, 1139, 1267, 2378], 'signalling.Nature.': [581], '2001;': [582], '411:': [583], '355-365Crossref': [584], 'PubMed': [585, 630, 689, 786, 806, 848, 868, 914, 961, 986, 1084, 1183, 1223, 1337, 1452, 1482, 1514, 1546, 1574, 1605, 1676, 1704, 1733, 1761, 1796, 1929, 1969, 1994, 2033, 2062, 2109, 2147, 2262, 2297, 2344, 2418, 2484, 2512, 2548, 2597, 2625, 2674, 2711, 2748, 2810, 2844, 3218, 3252, 3723, 3795, 3989], 'Scopus': [586, 631, 690, 787, 807, 849, 869, 915, 962, 987, 1049, 1085, 1156, 1184, 1224, 1284, 1338, 1453, 1483, 1515, 1547, 1575, 1606, 1677, 1705, 1734, 1762, 1797, 1930, 1970, 1995, 2034, 2063, 2110, 2148, 2263, 2298, 2345, 2395, 2419, 2485, 2513, 2549, 2598, 2626, 2675, 2712, 2749, 2811, 2845, 3219, 3253, 3724, 3796, 3990], '(3133)': [587], 'Google': [588, 633, 692, 789, 809, 851, 871, 917, 964, 989, 1051, 1087, 1158, 1186, 1226, 1286, 1340, 1455, 1485, 1517, 1549, 1577, 1608, 1679, 1707, 1736, 1764, 1799, 1932, 1972, 1997, 2036, 2065, 2112, 2150, 2265, 2300, 2347, 2397, 2421, 2487, 2515, 2551, 2600, 2628, 2677, 2714, 2751, 2813, 2847, 3221, 3255, 3726, 3798, 3992], 'Scholar).': [589, 810, 1088, 1227, 1341, 1800, 2151, 2422, 2552, 2629, 2752, 2848, 3222], 'At': [590], 'least': [591], 'one-third': [592], 'one-half': [594], 'human': [597, 684, 1787, 2694], 'proteins': [598], 'thought': [600, 1232], 'phosphorylated': [603, 976, 1299, 1325, 1433], 'at': [604, 942, 1831, 1910, 3016, 3022, 3049, 3064, 3105, 3122, 3179, 3299, 3320, 3411, 3667, 3950, 4074, 4142], 'some': [605, 3729, 3995], 'point': [606], '(2.Cohen': [607], 'regulation': [610], 'function': [613], 'by': [614, 838, 1405, 2133, 2216, 2326, 2411, 2449, 2661, 2697, 2738, 2915, 3090, 3245, 3340, 3687, 3960, 4129], 'multisite': [615], 'phosphorylation—a': [616], '25': [617, 3058, 3501], 'year': [618], 'update.Trends': [619], 'Biochem.': [620, 844], 'Sci.': [621], '2000;': [622, 2144], '25:': [623], '596-601Abstract': [624], 'Full': [625, 627, 1044, 1046, 1151, 1153, 1279, 1281, 1332, 1334, 1447, 1449, 2104, 2106, 2390, 2392, 2669, 2671, 2706, 2708, 2839, 2841], 'Text': [626, 628, 1045, 1047, 1152, 1154, 1280, 1282, 1333, 1335, 1448, 1450, 2105, 2107, 2391, 2393, 2670, 2672, 2707, 2709, 2840, 2842], 'PDF': [629, 1048, 1155, 1283, 1336, 1451, 2108, 2394, 2673, 2710, 2843], '(885)': [632], 'Scholar,': [634, 790, 1052, 1159, 1187, 1456, 1550, 1680, 1708, 1737, 1765, 1933, 1973, 1998, 2037, 2066, 2113, 2398, 2488, 2516, 2601, 2678, 2814], '3.Wilhelm': [635], 'M.': [636, 645, 667, 673, 880, 1165, 1658, 1774, 1943, 2232, 2273, 2654, 2680, 2784, 3198, 3694, 3766], 'Schlegl': [637], 'J.': [638, 669, 834, 1975, 2076, 2121, 2238, 2281, 2306, 2400, 2522, 2652, 2688, 2782, 3196, 3234, 3696, 3768], 'Hahne': [639], 'H.': [640, 655, 665, 876, 894, 1008, 1019, 1115, 1126, 1243, 1254, 1767, 1979, 2354, 2365, 2682, 3692, 3704, 3764, 3776], 'Moghaddas': [641], 'Gholami': [642], 'A.': [643, 675, 1552, 1583, 1682, 1945, 2070, 2234, 2275, 2490, 2603, 2780, 2822, 3700, 3772], 'Lieberenz': [644], 'Savitski': [646], 'M.M.': [647, 2047], 'Ziegler': [648], 'E.': [649, 3698, 3770], 'Butzmann': [650], 'L.': [651, 832, 1554, 1585, 1684, 2492, 2605], 'Gessulat': [652], 'S.': [653, 659, 773, 794, 878, 882, 1021, 1128, 1256, 1716, 1770, 1780, 2002, 2045, 2367, 3706, 3778], 'Marx': [654], 'Mathieson': [656], 'Lemeer': [658, 793], 'Schnatbaum': [660], 'K.': [661, 970, 2518, 2684, 2730], 'Reimer': [662], 'U.': [663], 'Wenschuh': [664], 'Mollenhauer': [666], 'Slotta-Huspenina': [668], 'Boese': [670], 'J.H.': [671], 'Bantscheff': [672], 'Gerstmair': [674], 'Faerber': [676], 'F.': [677, 2277, 2524, 2650], 'Kuster': [678], 'B.': [679, 792, 937, 1161, 1167, 1654, 1660, 1905, 2007, 2646, 2686], 'Mass-spectrometry-based': [680], 'draft': [681], 'proteome.Nature.': [685], '2014;': [686, 803, 1220, 2836], '509:': [687], '582-587Crossref': [688], '(1312)': [691], 'Scholar),': [693, 852, 872, 965, 990, 1287, 1486, 1518, 1578, 2266, 2301, 2348, 2715, 3727, 3993], 'hundreds': [695], 'different': [699, 1613, 2931], 'sites': [701, 2087], 'believed': [703], 'exist.': [705], 'Because': [706], 'this': [708, 1374, 2633, 2850, 2892], 'molecular': [709], 'complexity': [710, 1897], 'substoichiometric': [714], 'extent': [715], 'phosphorylation,': [717], 'specific': [718, 901], 'prior': [720, 2974], 'via': [723, 2639, 3361], 'liquid': [724, 746, 766, 2152, 2172, 2176, 2196, 2229, 2335], 'chromatography–tandem': [725, 733, 753, 2153, 2177], '(LC-MS/MS)': [728], '1The': [729, 749], 'abbreviations': [730, 750], 'used': [731, 751, 1884, 3163], 'are:LC-MS/MSliquid': [732, 752], 'spectrometryFDRfalse': [735, 755], 'discovery': [736, 756, 2157, 2181], 'rateIMACimmobilized': [737, 757], 'chromatographyACNacetonitrileFAformic': [741, 761], 'acidhSAXhydrophilic': [742, 762], 'exchangeHPLChigh-performance': [745, 765], 'chromatographyTiO2titanium': [747, 767], 'dioxide.': [748, 768, 2175, 2199], 'generally': [770, 1095, 1812], 'required': [771, 1813, 2769], '(4.Lemeer': [772], 'Heck': [774, 938, 1022, 1129, 1257, 1777, 1906, 2010, 2368, 3701, 3773], 'A.J.': [775, 939, 1907, 3702, 3774], 'phosphoproteomics': [777, 1315, 1476, 1508, 1540, 2019, 2055, 2290, 2478, 2534, 2591, 3983], 'data': [778, 2903, 2925], 'explosion.Curr.': [779], 'Opin.': [780], 'Chem.': [781, 864, 957, 982, 1219, 1478, 1510, 1542, 1925, 2480, 2593, 3985], 'Biol.': [782], '2009;': [783, 1730, 2341], '13:': [784, 2837], '414-420Crossref': [785], '(140)': [788], '5.Ruprecht': [791], 'Proteomic': [795], 'cancer.Expert': [800], 'Rev.': [801], 'Proteomics.': [802, 1041, 1148, 1276, 1328, 1443, 2100, 2387, 2544, 2666, 2702, 2835], '11:': [804, 2260], '259-267Crossref': [805], '(35)': [808], 'Most': [811, 1376], 'current': [812, 1343, 2758], 'such': [813, 1827], 'strategies': [814], 'exploit': [815], 'phosphate': [819], 'groups': [820], 'for': [821, 899, 978, 1071, 1236, 1317, 1361, 1567, 1598, 1697, 1752, 1818, 1885, 1958, 2053, 2253, 2288, 2407, 2505, 2618, 2971, 3019, 3052, 3061, 3108, 3119, 3164, 3202, 3210, 3241, 3317, 3806, 4071], 'metals': [822], 'carrier': [825], 'resins.': [826], 'These': [827], 'include': [829, 2226], 'Fe3+': [830], '(6.Andersson': [831], 'Porath': [833], 'Isolation': [835], 'phosphoproteins': [837], '(Fe3+)': [841], 'chromatography.Anal.': [843, 1218], '1986;': [845], '154:': [846], '250-254Crossref': [847], '(644)': [850], 'Ga3+': [853], '(7.Posewitz': [854], 'M.C.': [855], 'Tempst': [856], 'Immobilized': [858], 'gallium(III)': [859], 'phosphopeptides.Anal.': [863], '1999;': [865], '71:': [866], '2883-2892Crossref': [867], '(786)': [870], 'Zr4+': [874], '(8.Zhou': [875], 'Xu': [877], 'Ye': [879, 3693, 3765], 'Feng': [881], 'Pan': [883], 'C.': [884, 1772, 2041, 2236, 2279], 'Jiang': [885, 1978], 'X.': [886, 888, 1983], 'Li': [887], 'Han': [889], 'G.': [890], 'Fu': [891], 'Y.': [892, 1977, 1981, 2068, 3200], 'Zou': [893, 1018, 1125, 1253, 2364, 3703, 3775], 'Zirconium': [895], 'phosphonate-modified': [896], 'porous': [897], 'silicon': [898], 'highly': [900, 2657], 'capture': [902], 'MALDI-TOF': [906], 'MS': [907, 3395], 'analysis.J.': [908, 2256], 'Proteome': [909, 1728, 1756, 1791, 2028, 2057, 2257, 2292, 2339, 2805], 'Res.': [910, 1729, 1757, 1792, 2029, 2058, 2258, 2293, 2340, 2806], '2006;': [911, 983, 1991, 2059], '5:': [912, 2060], '2431-2437Crossref': [913], '(157)': [916], 'Scholar)': [918, 1609], '(IMAC);': [924], 'oxide': [926, 955, 1923], '(TiO2': [929], '(9.Pinkse': [930, 1898], 'M.W.': [931, 1899], 'Uitto': [932, 1900], 'P.M.': [933, 1901], 'Hilhorst': [934, 1902], 'M.J.': [935, 1903], 'Ooms': [936, 1904], 'Selective': [940, 971, 1908], 'isolation': [941, 1171, 1664, 1909], 'femtomole': [944, 1912], 'level': [945, 1913], 'proteolytic': [949, 1917], 'digests': [950, 1918, 3867], '2D-nanoLC-ESI-MS/MS': [952, 1920], 'titanium': [954, 1439, 1922, 2174, 2198, 3714, 3786], 'precolumns.Anal.': [956, 1924], '2004;': [958, 1926], '76:': [959, 1927], '3935-3943Crossref': [960, 1928], '(825)': [963, 1931], 'ZrO2': [966], '(10.Kweon': [967], 'H.K.': [968], 'Håkansson': [969], 'zirconium': [972], 'dioxide-based': [973], 'peptides': [977, 1300, 1434, 2127, 3209, 3293, 3310, 3587, 3920, 4103], 'spectrometric': [980], 'analysis.Anal.': [981], '78:': [984], '1743-1749Crossref': [985], '(470)': [988], 'others);': [992], 'recently': [995, 2456], 'introduced': [996, 1404], 'Ti-IMAC,': [997], 'which': [998, 1877], 'hybrid': [1001], 'other': [1004], 'two': [1005, 2982, 4107], '(11.Zhou': [1007, 1114, 1242, 2353], 'Low': [1009, 1116, 1244, 2355], 'T.Y.': [1010, 1117, 1245, 2356], 'Hennrich': [1011, 1118, 1246, 2357], 'M.L.': [1012, 1119, 1247, 2358], 'Toorn': [1013, 1120, 1248, 2359], 'H.v.': [1014, 1121, 1249, 2360], 'd.': [1015, 1122, 1250, 2361], 'Schwend': [1016, 1123, 1251, 2362], 'Mohammed': [1020, 1127, 1255, 1779, 2001, 2366, 3705, 3777], 'A.J.R.': [1023, 1130, 1258, 1778, 2011, 2369], 'Enhancing': [1024, 1131, 1259, 2370], 'putative': [1030, 1137, 1265, 2376], 'basophilic': [1031, 1138, 1238, 1266, 2377], 'substrates': [1033, 1140, 1268, 2379], 'Ti': [1035, 1142, 1270, 2381], '(IV)': [1036, 1143, 1271, 2382, 3715, 3787], 'based': [1037, 1144, 1272, 2383, 2859], 'enrichment.Mol.': [1039, 1146, 1274, 2385], 'Cell.': [1040, 1147, 1275, 1327, 1442, 2099, 2386, 2665, 2701, 2834], '2011;': [1042, 1149, 1277, 1479, 1511, 1543, 1571, 1602, 1701, 2294, 2388, 2481, 2509, 2594, 2622, 2667, 2807, 3986], '10Abstract': [1043, 1150, 1278, 2389, 2668], '(77)': [1050, 1157, 1285, 2396], '12.Lai': [1053], 'A.C.-Y.': [1054], 'Tsai': [1055], 'C.-F.': [1056, 1189], 'Hsu': [1057, 1190], 'C.-C.': [1058, 1191], 'Sun': [1059], 'Y.-N.': [1060], 'Chen': [1061, 1206, 2044], 'Y.-J.': [1062, 1207], 'Complementary': [1063], 'Fe3+-': [1064], 'Ti4+-immobilized': [1066], 'purification': [1072, 1364], 'acidic': [1074], 'basic': [1076], 'phosphopeptides.Rapid': [1077], 'Commun.': [1078, 1963], 'Mass': [1079, 1964], 'Spectrom.': [1080, 1965], '2012;': [1081, 2259, 2545, 2745], '26:': [1082], '2186-2194Crossref': [1083], '(45)': [1086], 'stated,': [1092], 'if': [1093], 'accepted,': [1096], 'capable': [1102], 'purifying': [1104, 1237], 'complementary': [1105, 1212], 'parts': [1106], 'characteristics': [1113, 2933], '13.Bodenmiller': [1160], 'Mueller': [1162, 1164, 1655, 1657], 'L.N.': [1163, 1656], 'Domon': [1166, 1659], 'Aebersold': [1168, 1661], 'R.': [1169, 1662, 1718, 2318, 2728], 'Reproducible': [1170, 1663], 'distinct,': [1173, 1666], 'overlapping': [1174, 1667], 'segments': [1175, 1668], 'phosphoproteome.Nat.': [1178, 1671], 'Methods.': [1179, 1672], '2007;': [1180, 1673, 3215], '4:': [1181, 1445, 1674, 2102], '231-237Crossref': [1182, 1675], '(505)': [1185, 1678], '14.Tsai': [1188], 'Hung': [1192], 'J.-N.': [1193], 'Wang': [1194, 2307], 'Y.-T.': [1195], 'Choong': [1196], 'W.-K.': [1197], 'Zeng': [1198, 1717, 2317], 'M.-Y.': [1199], 'Lin': [1200], 'P.-Y.': [1201], 'Hong': [1202], 'R.-W.': [1203], 'Sung': [1204], 'T.-Y.': [1205], 'Sequential': [1208], 'phosphoproteomic': [1209, 1754], 'through': [1211, 3344, 3879], 'metal-directed': [1213], '86:': [1221], '685-693Crossref': [1222], '(81)': [1225, 2299], 'For': [1228, 2958, 4145], 'example,': [1229], 'better': [1235], 'whereas': [1288], 'credited': [1291], 'efficient': [1295], 'multiply': [1298, 1324], '(15.Thingholm': [1301], 'T.E.': [1302, 1422], 'Jensen': [1303, 1423], 'O.N.': [1304, 1424], 'Robinson': [1305], 'P.J.': [1306], 'Larsen': [1307, 2527], 'M.R.': [1308, 1420, 2528], 'SIMAC': [1309], '(sequential': [1310], 'IMAC),': [1313], 'strategy': [1316, 2535], 'separation': [1320, 1751, 2213, 2321, 4147], 'monophosphorylated': [1322], 'peptides.Mol.': [1326], '2008;': [1329, 2030, 2415, 3249], '7:': [1330, 2031, 2746], '661-671Abstract': [1331], '(372)': [1339], 'consensus': [1344], 'field': [1347], 'no': [1350], 'method': [1351, 1566, 1597, 1696, 2504, 2617, 2893, 3033], 'superior': [1353], 'over': [1354, 2882, 3582, 3597, 3621, 3646], 'others': [1355], 'none': [1357], 'alone': [1358], 'sufficient': [1360, 2566], 'phosphoproteome,': [1367], 'view': [1369], 'challenge': [1372], 'study.': [1375], 'conducted': [1380], 'either': [1381], 'self-constructed': [1388], 'microcolumns': [1389], 'packed': [1390], 'pipette': [1392], 'tips.': [1393], 'Both': [1394], 'formats': [1395, 1828], 'have': [1396, 1622, 1881, 2565], 'been': [1397, 1861, 1882, 3312], 'considerable': [1402, 1624], 'variability': [1403], 'various': [1406], 'manual': [1407], 'steps': [1408, 4110], 'process.': [1411], 'Moreover,': [1412], 'differences': [1413], 'loading': [1415, 3837, 3864], 'conditions': [1416, 1810], 'including': [1417], 'additives': [1418], '(16.Larsen': [1419], 'Thingholm': [1421], 'Roepstorff': [1425], 'Jorgensen': [1427], 'T.J.': [1428], 'Highly': [1429, 2012], 'selective': [1430], 'peptide': [1436, 1469, 1501, 1533, 2212, 2471, 2584, 2789, 3165, 3976], 'mixtures': [1437, 2734], 'dioxide': [1440], 'microcolumns.Mol.': [1441], '2005;': [1444, 1966, 2101], '873-886Abstract': [1446], '(1333)': [1454], '17.Kettenbach': [1457], 'A.N.': [1458, 1490, 1522, 1776, 2460, 2573, 3965], 'Gerber': [1459, 1491, 1523, 2461, 2574, 3963, 3966], 'S.A.': [1460, 1492, 1524, 2462, 2575, 3967], 'Rapid': [1461, 1493, 1525, 2463, 2576, 2691, 3968], 'single-stage': [1464, 1496, 1528, 2466, 2579, 3971], 'mixtures:': [1470, 1502, 1534, 2472, 2585, 3977], 'application': [1471, 1503, 1535, 2473, 2586, 3363, 3978], 'general': [1473, 1505, 1537, 2475, 2588, 3980], 'phosphotyrosine-specific': [1475, 1507, 1539, 2477, 2590, 3982], 'experiments.Anal.': [1477, 1509, 1541, 2479, 2592, 3984], '83:': [1480, 1512, 1544, 2482, 2595, 3987], '7635-7644Crossref': [1481, 1513, 1545, 2483, 2596, 3988], '(129)': [1484, 1516, 1548, 2486, 2599, 3991], 'acid': [1487, 2166, 2190, 3493, 3526], 'concentration': [1488, 3027, 3083, 3301], '(17.Kettenbach': [1489, 1521, 2459, 2572, 3964], 'incubation': [1519, 4109], 'time': [1520, 2435], '18.Montoya': [1551, 1681, 2489, 2602], 'Beltran': [1553, 1584, 1683, 2491, 2604], 'Casado': [1555, 1586, 1685, 2493, 2606], 'Rodríguez-Prados': [1557, 1588, 1687, 2495, 2608], 'J.-C.': [1558, 1589, 1688, 2496, 2609], 'Cutillas': [1559, 1590, 1689, 2497, 2610], 'P.R.': [1560, 1591, 1690, 2498, 2611], 'Characterization': [1561, 1592, 1691, 2499, 2612], 'label-free': [1568, 1599, 1698, 2506, 2619], 'phosphoproteomics.Methods.': [1570, 1601, 1700, 2508, 2621], '54:': [1572, 1603, 1702, 2510, 2623], '370-378Crossref': [1573, 1604, 1703, 2511, 2624], '(94)': [1576, 1607, 1706, 2514, 2627], 'wash': [1580, 3509], 'volume': [1581], '(18.Montoya': [1582], 'lead': [1611], 'results.': [1614], 'One': [1615], 'prominent': [1616], 'parameter': [1617], 'effect': [1625], 'quality': [1629, 3733], 'bead': [1634, 4065], 'amount': [1635], 'digest': [1638, 3416], 'quantity,': [1639], 'finding': [1641], 'optimal': [1643], 'tradeoff': [1648], 'between': [1649], 'comprehensiveness': [1650], '(13.Bodenmiller': [1653], '19.Li': [1709], 'Q.R.': [1710], 'Ning': [1711], 'Z.B.': [1712], 'Tang': [1713], 'J.S.': [1714], 'Nie': [1715], 'Effect': [1719], 'peptide-to-TiO2': [1721], 'beads': [1722, 3262, 3307, 3328, 3681, 3848, 3998, 4038, 4082], 'selectivity.J.': [1727], '8:': [1731, 2342, 3721, 3793], '5375-5381Crossref': [1732], '(108)': [1735], '20.Yue': [1738], 'X.-S.': [1739], 'Hummon': [1740], 'A.B.': [1741], 'Combination': [1742], 'multistep': [1744], 'high-pH': [1748], 'reverse': [1749], 'phase': [1750, 2720], 'in-depth': [1753, 2254], 'profiling.J.': [1755], '2013;': [1758, 1793, 2703, 3720, 3792], '12:': [1759, 1794, 2704], '4176-4186Crossref': [1760], '(40)': [1763], '21.Zhou': [1766], 'Di': [1768], 'Palma': [1769], 'Preisinger': [1771], 'Peng': [1773], 'Polat': [1775], 'Toward': [1781], 'characterization': [1784], 'cancer': [1788], 'cell': [1789], 'phosphoproteome.J.': [1790], '260-271Crossref': [1795], '(297)': [1798], 'Consequently,': [1801], 'careful': [1802], 'priori': [1804], 'evaluation': [1805], 'optimization': [1807, 2962], 'case-by-case': [1816], 'basis': [1817], 'every': [1819], 'experimental': [1820], 'system.': [1821, 3534], 'Even': [1822], 'then,': [1823], 'use': [1825, 2240], 'comes': [1830], 'expense': [1833], 'intra-experimental': [1835], 'and,': [1836], 'even': [1837], 'so,': [1839], 'inter-experimental': [1840], 'accuracy.': [1841], 'means': [1844], 'overcome': [1846], 'issues,': [1848], 'direct': [1849, 2776], 'coupling': [1850], 'chromatographic': [1853, 2862, 4146], 'step': [1855, 2214, 2560], 'LC-MS/MS': [1858, 2640, 2662, 2777], 'system': [1859, 3456], 'applied.': [1862], 'Although': [1863, 2423], 'online': [1865, 1988, 2017, 2331], 'systems': [1866], 'increase': [1867, 3626], 'reproducibility,': [1868], 'sensitivity,': [1869], 'robustness,': [1871], 'limited': [1875, 1889], 'why': [1879], 'primarily': [1883], 'sample': [1890, 2887, 3183, 3467, 3756, 3807], 'amounts': [1891, 3184, 3808], 'samples': [1893, 3102, 3314], 'rather': [1895, 2928], 'low': [1896], '22.Ficarro': [1934], 'S.B.': [1935, 2117], 'Salomon': [1936], 'A.R.': [1937], 'Brill': [1938], 'L.M.': [1939], 'Mason': [1940], 'D.E.': [1941], 'Stettler-Gill': [1942], 'Brock': [1944], 'Peters': [1946], 'E.C.': [1947], 'Automated': [1948], 'chromatography/nano-liquid': [1952], 'chromatography/electrospray': [1953], 'ionization': [1954], 'platform': [1957], 'profiling': [1959], 'sites.Rapid': [1962], '19:': [1967], '57-71Crossref': [1968], '(89)': [1971], '23.Wang': [1974], 'Zhang': [1976, 2685], 'Cai': [1980], 'Qian': [1982], 'Phosphopeptide': [1984], 'detection': [1985], 'automated': [1987], 'IMAC-capillary': [1989], 'LC-ESI-MS/MS.Proteomics.': [1990], '6:': [1992], '404-411Crossref': [1993], '(46)': [1996], '24.Pinkse': [1999], 'M.W.H.': [2000], 'Gouw': [2003], 'J.W.': [2004], 'van': [2005], 'Breukelen': [2006], 'Vos': [2008], 'H.R.': [2009], 'robust,': [2013], 'automated,': [2014], 'sensitive': [2016, 2532, 2658], 'TiO2-based': [2018], 'study': [2022], 'endogenous': [2023], 'Drosophila': [2026], 'melanogaster.J.': [2027], '687-697Crossref': [2032], '(163)': [2035], '25.Ndassa': [2038], 'Y.M.': [2039], 'Orsi': [2040], 'Marto': [2042], 'J.A.': [2043], 'Ross': [2046, 2071], 'Improved': [2048], 'large-scale': [2054], 'applications.J.': [2056], '2789-2799Crossref': [2061], '(106)': [2064, 2713], '26.Zhang': [2067], 'Wolf-Yadlin': [2069], 'P.L.': [2072], 'Pappin': [2073], 'D.J.': [2074, 2820], 'Rush': [2075], 'Lauffenburger': [2077], 'D.A.': [2078], 'White': [2079, 2118], 'F.M.': [2080, 2119], 'Time-resolved': [2081], 'tyrosine': [2085], 'epidermal': [2090], 'growth': [2091], 'factor': [2092], 'receptor': [2093], 'signaling': [2094], 'network': [2095], 'reveals': [2096], 'dynamic': [2097, 2767], 'modules.Mol.': [2098], '1240-1250Abstract': [2103], '(474)': [2111], '27.Zarling': [2114], 'A.L.': [2115, 2818], 'Ficarro': [2116], 'Shabanowitz': [2120], 'Hunt': [2122], 'D.F.': [2123], 'Engelhard': [2124], 'V.H.': [2125], 'Phosphorylated': [2126], 'naturally': [2129], 'processed': [2130], 'presented': [2132], 'Major': [2134], 'Histocompatibility': [2135], 'Complex': [2136], 'Class': [2137], 'I': [2138], 'molecules': [2139], 'vivo.J.': [2141], 'Exp.': [2142], 'Med.': [2143], '192:': [2145], '1755-1762Crossref': [2146], '(167)': [2149], 'false': [2156, 2180], 'rate': [2158, 2182], 'acetonitrile': [2164, 2188], 'formic': [2165, 2189, 3492, 3525], 'high-performance': [2171, 2195], 'In': [2200, 2849, 4036], 'order': [2201, 2442], 'this,': [2204], 'workflows': [2207], 'employ': [2209], 'upstream': [2211], 'followed': [2215, 2448], 'each': [2220], 'fraction.': [2221], 'Examples': [2222], 'first-dimension': [2224], 'separations': [2225], 'interaction': [2228, 2244, 2270], '(28.Zarei': [2231], 'Sprenger': [2233, 2274], 'Gretzmeier': [2235, 2278], 'Dengjel': [2237, 2280], 'Combinatorial': [2239], 'electrostatic': [2242, 2267], 'repulsion-hydrophilic': [2243], '(ERLIC)': [2246], 'cation': [2249, 2351], '(SCX)': [2251], '4269-4276Crossref': [2261], '(32)': [2264], 'repulsion': [2268], '(29.Zarei': [2272], 'Metzger': [2276], 'Comparison': [2282], 'ERLIC–TiO2,': [2284], 'HILIC–TiO2,': [2285], 'SCX–TiO2': [2287], 'global': [2289, 2408, 2533, 3242], 'approaches.J.': [2291], '10:': [2295, 2808], '3474-3483Crossref': [2296], '(30.Dai': [2305], 'L.-S.': [2308], 'Wu': [2309, 2313], 'Y.-B.': [2310], 'Sheng': [2311], 'Q.-H.': [2312], 'J.-R.': [2314], 'Shieh': [2315], 'C.-H.': [2316], 'Fully': [2319], 'automatic': [2320], 'continuous': [2327], 'pH-gradient': [2328], 'coupled': [2332], 'reversed-phase': [2334], 'spectrometry.J.': [2338], '133-141Crossref': [2343], '(56)': [2346], '31.Villén': [2399, 3233], 'Gygi': [2401, 3235], 'S.P.': [2402, 3236], 'SCX/IMAC': [2404, 3238], 'approach': [2406, 2873, 3240], 'spectrometry.Nat.': [2413, 3247], 'Protoc.': [2414, 2744, 3214, 3248, 3719, 3791], '3:': [2416, 3250], '1630-1638Crossref': [2417, 3251], '(498)': [2420, 3254], 'powerful': [2424], 'frequently': [2426], 'after': [2429, 3674, 4057], 'fractionation': [2430, 2450], 'irreproducible': [2433], 'consuming.': [2436], 'consequence,': [2439], 'reversing': [2440], '(i.e.': [2443], 'first,': [2447], 'pool)': [2454], 'become': [2457], 'popular': [2458], '32.Engholm-Keller': [2517], 'Birck': [2519], 'Størling': [2521], 'Pociot': [2523], 'Mandrup-Poulsen': [2525], 'TiSH—a': [2529], 'robust': [2530, 2855], 'employing': [2536], 'combination': [2538, 2917], 'SIMAC,': [2541], 'HILIC.J.': [2543], '75:': [2546], '5749-5761Crossref': [2547], '(149)': [2550], 'This,': [2553], 'however,': [2554], 'requires': [2555], 'exquisite': [2563], 'selectivity,': [2564], 'allow': [2569], 'complete': [2570, 2772], 'Phosphopeptides': [2630], 'purified': [2631], 'way': [2634], 'directly': [2637, 3392], 'analyzed': [2638], '(33.Thakur': [2641], 'S.S.': [2642], 'Geiger': [2643], 'Chatterjee': [2645], 'Bandilla': [2647], 'Fröhlich': [2649], 'Cox': [2651, 2781], 'Mann': [2653, 2783, 3197], 'Deep': [2655], 'proteome': [2659, 2695], 'coverage': [2660], 'without': [2663], 'prefractionation.Mol.': [2664], '(270)': [2676], '34.Pirmoradian': [2679], 'Budamgunta': [2681], 'Chingin': [2683], 'Astorga-Wells': [2687], 'Zubarev': [2689], 'R.A.': [2690], 'deep': [2693], 'single-dimension': [2698], 'shotgun': [2699, 2794], 'proteomics.Mol.': [2700], '3330-3338Abstract': [2705], 'typically': [2716], 'shallow': [2718], 'reversed': [2719], 'LC': [2721], 'gradients': [2722], '(35.Köcher': [2723], 'Pichler': [2725], 'Swart': [2727], 'Mechtler': [2729], 'Analysis': [2731], 'whole-cell': [2736], 'extracts': [2737], 'single-run': [2739], 'nanoLC-MS/MS': [2740], 'ultralong': [2742], 'gradients.Nat.': [2743], '882-890Crossref': [2747], '(90)': [2750], 'it': [2754, 3530], 'turns': [2755], 'spectrometers': [2760], 'still': [2761], 'lack': [2762], 'scan': [2764], 'speed': [2765], 'reach': [2771], '(phospho)peptide': [2773], 'sampling': [2774], '(36.Michalski': [2779], 'More': [2785], '100,000': [2787], 'detectable': [2788], 'species': [2790], 'elute': [2791], 'single': [2793], 'proteomics': [2795, 3211], 'runs': [2796], 'majority': [2799], 'inaccessible': [2801], 'LC-MS/MS.J.': [2804], '1785-1793Crossref': [2809], '(476)': [2812], '37.Hebert': [2815], 'A.S.': [2816], 'Richards': [2817], 'Bailey': [2819], 'Ulbrich': [2821], 'Coughlin': [2823], 'E.E.': [2824], 'Westphall': [2825], 'M.S.': [2826], 'Coon': [2827], 'J.J.': [2828], 'one': [2830], 'hour': [2831], 'yeast': [2832], 'proteome.Mol.': [2833], '339-347Abstract': [2838], '(411)': [2846], 'article,': [2851], 'describe': [2853], 'flexible': [2857], 'commercially': [2868], 'available': [2869], 'column.': [2871], 'This': [2872], 'offers': [2874], 'scales': [2881], 'quantities.': [2888], 'show': [2890], 'tested,': [2900], 'our': [2902], 'argue': [2904], 'apparent': [2907], 'three': [2911, 3678], 'caused': [2914], 'inadequacies,': [2920], 'speed,': [2927], 'phosphopeptides.': [2935], 'Human': [2936], 'epidermoid': [2937], 'A431': [2938], 'cells': [2939, 2964, 2979], 'were': [2940, 2965, 2980, 3126, 3162, 3174, 3190, 3226, 3268, 3294, 3315, 3329, 3376, 3382, 3406, 3538, 3588, 3654, 3682, 3824, 3849, 3868, 3888, 3921, 3945, 4006, 4039, 4048, 4083, 4104], 'grown': [2941], "Iscove's": [2943], 'modified': [2944, 3805], "Dulbecco's": [2945], 'medium': [2946], 'supplemented': [2947], '10%': [2949, 3928], '(v/v)': [2950, 3429, 3617, 3898, 3901, 3914, 3917, 3929, 3936, 3939, 4121], 'fetal': [2951], 'bovine': [2952], 'serum': [2953], '1%': [2955, 3356, 3388], 'antibiotic/antimycotic': [2956], 'solution.': [2957, 4101], 'experiments,': [2963], 'treated': [2966], '1': [2968, 3020, 3053, 3144, 3273, 3303, 3318, 3818, 3833, 3862, 3871, 3896, 4010, 4023, 4072, 4097], 'mm': [2969, 2994, 3047, 3059, 3077, 3279, 3369, 3441, 3446, 3491, 3502, 3505, 3524, 3554, 3904, 4118], 'pervanadate': [2970], 'min': [2973, 3063], 'lysis.': [2976], 'After': [2977, 3305, 3567, 3624, 3882], 'harvesting,': [2978], 'washed': [2981, 3269, 3339, 3383, 3589, 3889, 4007, 4084], 'times': [2983, 3271, 4021, 4086, 4095], 'ice-cold': [2985], 'PBS': [2986], 'lysed': [2988], '8': [2990], 'm': [2991, 4029], 'urea,': [2992], '40': [2993, 3076, 3353, 3385, 3398], 'Tris/HCl': [2995, 3078], '(pH': [2996], '7.6),': [2997], '1×': [2998, 3007], 'EDTA-free': [2999], 'protease': [3000], 'inhibitor': [3001, 3009], 'mixture': [3002, 3010, 3072, 4068], '(Complete': [3003], 'Mini,': [3004], 'Roche),': [3005], 'phosphatase': [3008], '(Sigma).': [3011], 'lysate': [3013], 'centrifuged': [3015], '20,000': [3017], 'rpm': [3018], 'h': [3021, 3054, 3319, 4073], '4': [3023, 3109], '°C.': [3024, 3124, 3181, 3413, 3669, 3952, 4144], 'determined': [3029], 'Bradford': [3032], '(Coomassie': [3034], '(Bradford)': [3035], 'Assay': [3037], 'Kit,': [3038], 'Thermo': [3039, 3449], 'Scientific).': [3040], 'supernatant': [3042, 4044, 4126], 'reduced': [3044], '10': [3046, 3557, 3583, 3891, 3907], 'DTT': [3048], '56': [3050], '°C': [3051, 3107], 'alkylated': [3056], 'iodoacetamide': [3060], '45': [3062], 'room': [3065, 3321, 4075], 'temperature': [3066, 3322, 4076], 'dark.': [3069], 'diluted': [3074], 'final': [3081], 'urea': [3082], '1.6': [3085], 'm.': [3086], 'induced': [3088], 'digestion': [3089, 3121], 'adding': [3091], 'sequencing-grade': [3092], 'trypsin': [3093, 3116, 3136], '(Promega,': [3094], 'Mannheim,': [3095], 'Germany;': [3096, 3151], '1:100': [3097, 3113], 'enzyme:substrate': [3098], 'ratio)': [3099], 'allowing': [3101], 'incubate': [3104], '37': [3106, 3123], 'h.': [3110], 'Subsequently': [3111], 'another': [3112], 'quantity': [3114], 'added': [3118], 'overnight': [3120], 'Samples': [3125], 'acidified': [3127], 'TFA': [3129], 'pH': [3131, 3371, 4123], '2': [3132, 3912], 'stop': [3134], 'activity.': [3137], 'SepPack': [3138], '(C18': [3140, 3814], 'cartridges,': [3141, 3815], 'Sep-Pak': [3142, 3812, 3816], 'Vac,': [3143, 3817], 'cc': [3145, 3819], '(50': [3146, 3820, 4117], 'mg),': [3147, 3821], 'Waters': [3148, 3822], 'Corp.,': [3149], 'Eschborn,': [3150], 'solvent': [3152, 3156, 3277, 3289, 3298, 3350, 3424, 3593, 3614, 3629, 3642, 3838, 3875, 4014, 4027, 4092, 4116], 'A,': [3153], '0.07%': [3154, 3158, 3428], 'TFA;': [3155], 'B,': [3157], 'TFA,': [3159, 3902], '50%': [3160, 3285, 4032], 'ACN)': [3161], 'desalting': [3166], 'according': [3167, 3475, 3656], "manufacturer's": [3170, 3478], 'instructions,': [3171], 'eluates': [3173], 'dried': [3175, 3407, 3663, 3946, 4138], 'down': [3176, 3408, 3947, 4041], 'stored': [3178, 3410, 3666, 3949, 4141], '−80': [3180, 3412, 3668, 3951, 4143], 'Smaller': [3182], 'up': [3186, 3751], 'desalted': [3191, 3415], 'as': [3192, 3229, 3684, 3759, 3958], 'described': [3193, 3230, 3686, 3760, 3959], 'elsewhere': [3194], '(38.Rappsilber': [3195], 'Ishihama': [3199], 'Protocol': [3201], 'micro-purification,': [3203], 'enrichment,': [3204], 'pre-fractionation': [3205], 'storage': [3207], 'StageTips.Nat.': [3213], '2:': [3216], '1896-1906Crossref': [3217], '(2570)': [3220], 'essentially': [3227], 'performed': [3228, 3758, 3957], 'Ref.': [3232, 3762], 'Scholar.': [3256], 'Briefly,': [3257, 3480], 'μl': [3259, 3346, 3354, 3366, 3386, 3399, 4113, 4134], '(PhosSelect': [3263], 'iron': [3264, 3474, 3512, 3565], 'gel,': [3266], 'Sigma)': [3267], 'four': [3270, 4020, 4085], 'ml': [3274, 3421, 3543, 3551, 3558, 3834, 3863, 3872, 3892, 3908, 3926, 3933, 4011, 4024, 4052, 4062, 4089, 4098], '(25': [3278], 'FA,': [3280, 4137], '40%': [3281], 'ACN),': [3282], 'slurry': [3286], 'prepared.': [3291], 'Dried-down': [3292, 3866], 'resuspended': [3295], 'μg/μl.': [3304], 'dissolved': [3309, 3869, 4049], 'had': [3311], 'combined,': [3313], 'incubated': [3316, 4070], 'vigorous': [3324, 4078], 'shaking.': [3325, 4079], 'Subsequently,': [3326, 3843, 4080], 'transferred': [3330], 'top': [3332, 3852], 'previously': [3334, 3685], 'equilibrated': [3335, 3858, 4064], 'C18': [3336, 3380, 3855], 'StageTips': [3337], 'being': [3341], 'sequentially': [3342, 3923], 'passed': [3343, 3878], '50': [3345, 3440, 3445, 3553, 3844], '(twice)': [3351], 'FA.': [3357, 3404, 3940], 'Elution': [3358], 'achieved': [3360, 3604], '70': [3365], '500': [3368], 'K2HPO4,': [3370], '7': [3372], '(twice).': [3373], 'Peptides': [3374, 4047], 'retained': [3377], 'FA': [3389], 'eluted': [3391, 3922, 4105], 'plate': [3396], '60%': [3401], 'ACN,': [3402, 3427, 3840, 3899, 3915, 3937, 4016, 4033], '0.1%': [3403, 3916, 4017, 4034], 'Eluates': [3405], 'reconstituted': [3418], '0.5': [3420, 4051], 'A': [3425, 3594, 3643, 3736], '(30%': [3426], 'TFA)': [3430, 4018], 'loaded': [3432, 3579, 3850], 'onto': [3433], 'analytical': [3435], '(4': [3438], '×': [3439, 3444, 3861], '9': [3443], 'ProPac': [3447], 'IMAC-10,': [3448], 'Fisher': [3450], 'Scientific)': [3451], 'connected': [3452], '(ÄKTA': [3457], 'Explorer': [3458], 'FPLC': [3459], 'system,': [3460], 'Amersham': [3461], 'Biosciences': [3462], 'Pharmacia)': [3463], '1-ml': [3466], 'loop.': [3468], 'charged': [3472, 3495], 'instructions.': [3479], 'rinsed': [3484], '3': [3486, 3497], 'volumes': [3488, 3499, 3521], '20': [3490, 3519, 3523, 3542, 3550], 'FeCl3,': [3503], 'acetic': [3506], 'acid.': [3507], 'unbound': [3511, 3586], 'ions,': [3513], 'flushed': [3515, 3539, 3831], 'subsequently': [3528], 'removed': [3529], 'lines': [3537], 'first': [3540], 'double-deionized': [3545, 3560], 'H2O': [3546, 3561], 'then': [3548], 'EDTA': [3555], 'remove': [3563], 'remaining': [3564], 'ions.': [3566], 'reconnection': [3569], 'baseline': [3571], 'equilibration,': [3572], 'gradient': [3574, 3608], 'started.': [3576], 'Sample': [3577], '(0.1': [3580], 'ml/min': [3581, 3596, 3620, 3645], 'min),': [3584], '(0.3': [3595], '16': [3598], 'min).': [3599, 3623, 3648], 'Subsequent': [3600], 'linear': [3607], '0%': [3610], '45%': [3612], 'B': [3615, 3630], '(0.5%': [3616], 'NH4OH)': [3618], '(0.2': [3619], '60': [3622], '100%': [3628, 4136], '5-min': [3633], 'holding': [3634], 'step,': [3635], 're-equilibrated': [3639], '(0.5': [3644], '30': [3647, 4133], 'Flow-through': [3649, 3941], 'fraction': [3653], 'collected': [3655], 'UV': [3659], 'signal': [3660], '(280': [3661], 'nm),': [3662], 'down,': [3664, 4139], 'recharged': [3673], 'maximum': [3676], 'enrichments.': [3679], 'synthesized': [3683], 'Zhou': [3688], 'et': [3689], 'al.': [3690], '(39.Zhou': [3691], 'Dong': [3695, 3767], 'Corradini': [3697, 3769], 'Cristobal': [3699, 3771], 'Robust': [3707, 3779], 'monodisperse': [3711, 3783], 'microsphere-based': [3712, 3784], 'chromatography.Nat.': [3718, 3790], '461-480Crossref': [3722, 3794], '(243)': [3725, 3797], 'modifications': [3730], 'additional': [3732], 'control': [3734], 'steps.': [3735], 'detailed': [3737], 'description': [3738], 'supplemental': [3744], '"Materials': [3745], 'Methods"': [3747], 'section.': [3748], 'Enrichment': [3749], '250': [3753, 3810], '39.Zhou': [3763], 'Scholar;': [3799], 'protocol': [3803], 'exceeding': [3809], 'μg.': [3811], 'cartridges': [3813], 'Corp.)': [3823], 'attached': [3825], 'vacuum': [3828], 'manifold': [3829], '(80%': [3839], '6%': [3841], 'TFA).': [3842, 3918, 4035], 'solvent.': [3865], 'slowly': [3877], 'cartridges.': [3881], 'reapplication': [3883], 'flow-through,': [3886], 'washing': [3894, 3910, 4013, 4100], 'solution': [3895, 3911], '(50%': [3897, 3913, 4015], '0.5%': [3900, 4120], '200': [3903, 4112], 'NaCl)': [3905], 'Bound': [3919, 4102], '0.8': [3925], 'NH4OH': [3930], '0.2': [3932, 4088], '80%': [3935], '2%': [3938], 'fractions': [3944], 'Kettenbach': [3961], 'modifications.': [3996], '(5': [3999], 'μm,': [4000], 'GL': [4001], 'Sciences': [4002], 'Inc.,': [4003], 'Mainz,': [4004], 'Germany)': [4005], 'twice': [4008], '(2': [4028], 'lactic': [4030], 'acid,': [4031], 'between,': [4037], 'spun': [4040], 'discarded.': [4046], 'solvent,': [4055], 'addition': [4059, 4131], '0.25': [4061], 'slurry,': [4066], 'five': [4094], '10-min': [4108], 'KH2PO4,': [4119], 'NH4OH,': [4122], '11.3).': [4124], 'quenched': [4128]}, 'cited_by_api_url': 'https://api.openalex.org/works?filter=cites:W2128134586', 'counts_by_year': [{'year': 2024, 'cited_by_count': 7}, {'year': 2023, 'cited_by_count': 8}, {'year': 2022, 'cited_by_count': 15}, {'year': 2021, 'cited_by_count': 10}, {'year': 2020, 'cited_by_count': 20}, {'year': 2019, 'cited_by_count': 14}, {'year': 2018, 'cited_by_count': 12}, {'year': 2017, 'cited_by_count': 18}, {'year': 2016, 'cited_by_count': 11}, {'year': 2015, 'cited_by_count': 9}], 'updated_date': '2024-12-26T22:19:18.423716', 'created_date': '2016-06-24'}
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