Proteomic analysis of mouse astrocytes and their secretome by a combination of FASP and StageTip‐based,high pH,reversed‐phase fractionation |
| |
| Authors: | Dohyun Han Jonghwa Jin Jongmin Woo Hophil Min Youngsoo Kim |
| |
| Institution: | 1. Department of Biomedical Sciences, Seoul National University College of Medicine, , Seoul, Korea;2. Department of Biomedical Engineering, Seoul National University College of Medicine, , Seoul, Korea;3. Institute of Medical & Biological Engineering, Medical Research Center, Seoul National University College of Medicine, , Seoul, Korea |
| |
| Abstract: | Astrocytes are the most abundant cells in the CNS, but their function remains largely unknown. Characterization of the whole‐cell proteome and secretome in astrocytes would facilitate the study of their functions in various neurodegenerative diseases and astrocyte–neuron communication. To build a reference proteome, we established a C8‐D1A astrocyte proteome to a depth of 7265 unique protein groups using a novel strategy that combined two‐step digestion, filter‐aided sample preparation, StageTip‐based high pH fractionation, and high‐resolution MS. Nearly, 6000 unique protein groups were identified from conditioned media of astrocyte cultures, constituting the largest astrocyte secretome that has been reported. High‐confidence whole‐cell proteomes and secretomes are valuable resources in studying astrocyte function by label‐free quantitation and bioinformatics analysis. All MS data have been deposited in the ProteomeXchange with identifier PXD000501 ( http://proteomecentral.proteomexchange.org/dataset/PXD000501 ). |
| |
| Keywords: | Astrocyte High pH fractionation LC‐MS/MS Proteome profile Secretome Technology |
|
|
