Isolated proteins were analyzed and identified using LC–MS Repre

Isolated proteins were analyzed and identified using LC–MS. Representative proteins are shown in Table 2. Fig. 1 a PAGE of IP samples using anti-human IgA antibody-conjugated Dynabeads. ‘M’ represents the molecular weight markers. IP samples were derived from urine of IgAN patients (lanes 1 and 2) and a healthy control (lane 3). b PAGE of IP samples using BSA blocking Dynabeads. ‘M’ represents the molecular weight markers. IP samples were derived from urine of IgAN patients

(lanes 1 and 2) and a healthy control (lane 3) Table 2 Summary of the LC–MS analysis result of the protein collected from the urine of IgAN patients and healthy donors by IP method using anti-IgA conjugated beads and mTOR inhibitor cancer BSA beads Beads: anti-IgA conjugated beads BSA beads Disease: IgAN Other kidney diseases IgAN Sample no: 1 2 3 4 10 11 12 5 6 7 8 9 1 2   ID Protein name                             Cell component or other gi|340166 Uromodulin 3 3   1 3   1 1 1           gi68838 Aquaporin               1 1           gi|7331218 Keratin 1 2 2 2     1       2 1 2 1 2 gi|34073 Cytokeratin 4 (408 AA) 1 1   1       1             gi186629 Keratin 10           1       1   1     gi|34033

HMPL-504 concentration Keratin 13 1 1                         gi177139 Keratin 14       1   1         1 1     gi186685 Keratin 16           1 1       1       gi34081 Keratin 17                   1         Serum protein gi|4557871 Transferrin 14 14     1           1   1   gi|28592 Serum albumin 3 45 6 2 4   3 2 1   5 3   3 gi|4557385 Complement component 3 (C3) 1 3                     1   gi|306882 Haptoglobin precursor 2 3                         gi|72059 Leucine-rich alpha-2-glycoprotein 1 2                     2   gi177827 Alpha-1-antitrypsin       1 2 2 2   1   2       gi45067732 S100 calcium-binding protein A9         1 2       selleck products           gi|493852 Hemoglobin 5 1       1 1           8 2 gi|224053 Macroglobulin alpha2 1 2                         Antibody component

gi|223099 IgA alpha1 Bur 2 1                         gi|223335 Ig kappa L I Den 1 1                         gi|229528 Protein Len, Bence-Jones 2 3                     1   gi33700 Ig lambda light chain 1 2 1         1     1 1     gi9857759 IgG4 heavy chain                     1       gi229526 Protein Rei, Bence-Jones     3               5         Ig kappa light chain 3 3                 2         Ig heavy chain 2 4 2               1       Urine samples were from IgAN patients (1, 2, 3, 4, 10, 11, 12), amyloidosis (5), SLE (6), DMN (7, 8), and MCNS (9). The numbers in the P005091 clinical trial column show the identified number of fragments by LC–MS analysis Western blot analysis of the IgA–uromodulin complex The results of LC–MS analysis were confirmed by Western blot (WB) analysis using antibodies against the identified proteins. Figure 2 is an example of the analysis of uromodulin. Uromodulin was strongly positive in the urine samples of seven IgAN patients.

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