| Metadata |
| datasetIdentifier | PASS01173 |
| datasetType | SRM |
| submitter | Laura Mourino-Alvarez <lmourino@sescam.jccm.es> |
| submitter_organization | Hospital Nacional de Parapléjicos (SESCAM) |
| lab_head_full_name | Maria G Barderas |
| lab_head_email | megonzalezb@sescam.jccm.es |
| lab_head_organization | Hospital Nacional de Parapléjicos (SESCAM) |
| lab_head_country | Spain |
| datasetTag | HuPlasma_TPMLDHBTERA |
| datasetTitle | Crude Human Plasma_Applied Biosystems 4000 QTRAP LC/MS/MS System_90 min run_TPM-1, LDHB, TERA |
| publicReleaseDate | 2018-09-30 00:00:00 |
| finalizedDate | |
| summary | Peripheral blood samples were collected from control subjects and patients with severe aortic stenosis obtained from subjects who underwent scheduled AV replacement. |
| contributors | Laura Mourino-Alvarez
Maria G Barderas |
| publication | Mourino-Alvarez, L et al. A comprehensive study of calcific aortic stenosis: from rabbit to human samples. Dis Model Mech. 2018 May 10. pii: dmm.033423. doi: 10.1242/dmm.033423.
|
| growth | |
| treatment | |
| extraction | Proteins were reduced and alkylated by incubating with 100 mM DTT and 550 mM iodoacetamide in 50 mM ammonium bicarbonate, respectively. |
| separation | |
| digestion | Proteins were digested in 50 mM ammonium bicarbonate, 15% acetonitrile with sequencing grade modified porcine trypsin. After overnight digestion, 2% formic acid was added and samples were cleaned with Pep‐Clean spin columns (Pierce). Tryptic digests were dried in a speedvac and resuspended in 2% ACN, 2% formic acid prior to MS analysis. |
| acquisition | Three replicate injections were made for each sample using mobile phase A (2% ACN/98% water, 0.1% formic acid [FA]) with a flow rate of 10μl L/min for 5min. Peptides were loaded onto a μ‐Precolumn Cartridge (Acclaim Pep Map 100 C18, 5 μm, 100Å; 300 μm i.d.× 5mm, LC Packings) to preconcentrate and desalt samples. Reverse-phase liquid chromatography was achieved on a C18 column (Onyx Monolithic C18, 150 × 0.1mm i.d., Phenomenex) in a gradient of phase A and phase B (98% ACN/2% water, 0.1% FA). Peptides were eluted at a flow rate of 900 nlL/min in the following steps: 5–45% B for 60 min, 45–95% B for 1 min and finally 95% B for 4 min. The mass spectrometer was set to operate in positive ion mode with ion spray voltage of 2800V and a nanoflow interface heater temperature of 80ºC. Source gas 1 and curtain gas were both set to 20, and nitrogen was applied as both curtain and collision gases. |
| informatics | |
| instruments | Applied Biosystems 4000 QTRAP LC/MS/MS System |
| species | Human |
| massModifications | none |
