Project description:The absolute protein amounts for the selected yeast ribosomal proteins (RPs) were determined using the standard peptides and TMT labelling. The range of the standard tryptic peptides were selected and synthesized based on the preliminary data-dependent experiments. The synthetic peptides were mixed and four aliquots containing different amounts of the peptides were labelled each with a different TMT labelling reagent. The yeast cell lysate was digested and labelled in parallel. The labelled cellular and standard samples were combined, and the absolute peptide amounts in the lysate were calculated by comparison to the known amounts in the standard TMT channels.
Project description:Hela cells were treated with a mixture of amino acids and other small molecules, called active mixture for various time periods. The aim was to identify the differentially expressed genes that are upregulated or downregulated upon treatment at different time points.
Project description:Protein extracts of Saccharomyces cerevisiae CEN.PK113-7D cultivated in chemostats under different conditions. Representative samples containing aliquots of all conditions were spiked with UPS2 standard (Sigma) to estimate absolute values in fmol. The conditions for Saccharomyces cerevisiae CEN.PK113-7D are: T2- Standard condition : 30°C, pH 5.5 T3- High temperature: 36°C, pH 5.5 T4- Low pH: 30°C, pH 3.5 T5- Osmotic stress : 30°C, pH 5.5, 1M KCl T6- Anaerobic condition Furthermore, representative samples pooling aliquots of each condition are indicated as "bulk" samples. These samples were spiked with UPS proteins. A validation step was carried out by spiking 4 external proteins at known concentrations within the yeast and UPS proteins mixture
Project description:Comparing the performance of methylamine and hydroxylamine on phospho-peptide analysis, particularly in relation to the identification of more complex dose-dependent patterns.