Shotgun Proteomics Analysis of the Osteoblasts Differentiation of Rat Bone Marrow Stem Cells

摘要

The combined application of SDS-PAGE and RP-HPLC separation of peptides provided the identification of proteins limited by a specific range of MW decreased chances for false positive protein identification and increased the number of low copies proteins identified in this work in contrast to shotgun proteomics using combination of ion exchange/reverse phase HPLC for total digestion of protein cell extracts. An average of 23 protein per fraction of SDS-PAGE (6 fractions in total) were identified. This strategy could be more effective for proteomic analysis of low amount of proteins than 2D gel electrophoresis. We conclude that the acquired protein list of rat BM-MSCs by shotgun proteomics in this study can be integrated into inventory, which may facilitate the identification of the proteomic pattern as well as changes in activated or suppressed pathways occurrin g during differentiation and other experimental conditions.