Project description:Nano-flow chromatography-tandem mass spectrometer (nLC-MS/MS) is a popular choice in proteome research as it provided high-sensitivity analysis with minimal sample usage, but its quantitative proteomics applications become restricted by low analytical throughput, and low robustness. To circumvent this limitation, we describe the performance of the new Vacuum Insulated Probe Heated ElectroSpray Ionization source (VIP-HESI) coupled to the Bruker timsTOF mass spectrometer and show it enhances micro-spray flow rate chromatography signals in comparison to nanospray source conditions using the CaptiveSpray (CS) and ElectroSpray Ionization (ESI) sources. In addition, using a 0.5 x 200mm and 1 × 150 mm columns, achieved excellent reproducibility of chromatographic retention time, coefficient of variation, and precision quantification using data from un-depleted mouse plasma, HeLa and K562 digest peptide samples.
Project description:Analysis of the ovarian cancer cell line OVCAR-5. A standard trypsin digest was carried out on the OVCAR-5 cell lysates which were then analysed in the un-fractionated and fractionated forms. Fractionation was completed using a peptide IEF separation method. All samples were analysed by nano-LC-ESI-MS/MS using a QTOF.
Project description:To identify proteotypic peptides, HeLa cells digest labeled with mTRAQdelta0 were extensively fractionated and analyzed by LC-MS/MS.
Project description:1) AJS-ESI source comparison of 2, 10 and 20ug of HeLa digest
2) Label-free analysis of 11-week NPC1 null mice and control for unfractionated cerebellum and cerebral cortex
3) Label-free analysis of 11-week NPC1 null mice and control for fractionated cerebellum
Project description:This technical dataset contains LC-MS runs of HeLa protein digest analyzed with a common shotgun proteomics pipeline based on MSGF+ and Percolator and DirectMS1 pipeline.
Project description:Comparison of in-solution and single-pot, solid-phase-enchanced sample preparation (SP3) sample processing workflows on HeLa cell samples, using different lysis buffers based on SDS and guanidine hydrochloride, for optimal analysis by liquid chromatography mass spectrometry (LC-MS)