Selected reaction monitoring based analysis of proteins in liver tissue of Labeo rohita

Bottom-up proteomics analysis relies on efficient protein digestion for performing mass spectrometric analysis. Optimisation of protein solubilisation and digestion strategies is a key step for obtaining reliable proteomic data. In this study, two different solubilisation buffers (SDS and Urea) and three digestion methods (FASP, S-Trap and in-solution,) were applied for the proteomic analysis of liver tissue of Labeo rohita. Label free quantification analysis was performed to analyse the similarities and differences in the results obtained in each method. Based on the discovery data, few proteins were selected for targeted data acquisition. These proteins were chosen on the basis of highest peptide spectral matches in different experimental conditions. The aim was to validate the trends in abundance of proteins in different experimental conditions.