| Metadata |
| datasetIdentifier | PASS00661 |
| datasetType | SRM |
| submitter | yunki yau <yunki.yau@outlook.com> |
| submitter_organization | Bioanalytical Mass Spectrometry Facility, Mark Wainwright Analytical Centre, The University of New South Wales |
| lab_head_full_name | Mark Raftery |
| lab_head_email | m.raftery@unsw.edu.au |
| lab_head_organization | Bioanalytical Mass Spectrometry Facility, Mark Wainwright Analytical Centre, The University of New South Wales |
| lab_head_country | AUSTRALIA |
| datasetTag | SerumFABP5 |
| datasetTitle | RPD Calibration curve for human serum Fatty Acid-Binding Protein 5 (FABP5) proteotypic peptide assay. |
| publicReleaseDate | 2015-06-01 00:00:00 |
| finalizedDate | 2017-03-06 16:35:22 |
| summary | This dataset is the RPD calibration curve data for a tier 2 MRM human serological assay (NCI-Clinical Proteomic Tumor Analysis Consortium (CPTAC)) for a proteotypic peptide of Fatty Acid-Binding Protein 5. |
| contributors | Yunki Yau, Rupert Leong, Aviv Pudipeddi, Diane Redmond, Valerie Wasinger |
| publication | Yau YY, Leong RWL, Pudipeddi A, Redmond D, Wasinger VC, Serological Epithelial Component Proteins Identify Intestinal Complications in Crohn's disease, Molecular & Cellular Proteomics, submitted |
| growth | N/A |
| treatment | 20mL of peripheral blood were drawn from consenting subjects in two 20mL SST tubes. Serum was separated from blood cells by centrifugation at 1400 rpm and frozen at -80°C prior to analysis. All participants gave written informed consent. |
| extraction | serum samples desalted with C18 stagetip columns. |
| separation | Samples were concentrated and desalted onto a micro C18 precolumn (500 µm x 2 mm, Michrom Bioresources, USA) with H2O:CH3CN (98:2, 0.05 % v/v TFA) at 15 l/minute. After 4 min washing the pre-column was automatically switched (Valco10 port valve, Houston, USA) into line with a fritless-nano column manufactured according to Gatlin et al., (1998).(22) Peptides were eluted using a linear gradient of H2O:CH3CN (98:2, 0.1 % (v/v) FA) to H2O:CH3CN (36:64, 0.1 % (v/v) FA) at ~300 nL/min over 40 min. The pre-column was connected via a fused silica capillary (25 cm, 25 µm) to a low volume tee (Upchurch Scientific, USA) and introduced into the 4000QTRAP mass spectrometer. |
| digestion | 2μl of serum (equating to 57µg ± 7% of protein) were trypsin digested at an approximate 1:100 enzyme-to-protein ratio. |
| acquisition | Samples were analyzed in positive ion mode with an ion spray voltage of 2.4 kV, curtain gas flow of 20 and nebulizing gas flow of 5. For MRM analysis, quadrupoles were operated in unit resolution, and the dwell time was 66.2ms. All samples were made up to 10µL and analyzed in 1µL injections. |
| informatics | The transition list was developed in Skyline SRM environment v1.4 (MacCoss Lab, UW) using MS/MS information from the SRMAtlas human library (Institute for Systems Biology, WA, USA). Raw .WIFF files were imported into Skyline SRM Environment for manual inspection and peak area-ratio calculation. |
| instruments | AB SCIEX 4000 QTrap |
| species | Human |
| massModifications | R+10.008269 |
