| JPST000161 | |
| jPOST ID | JPST000161 |
| PXID | PXD005096 |
| Project title | Microproteomics application for the limited number of cells |
| Keywords | THP-1, shotgun proteomics, Cell-sorting, microproteomics |
| Description | Mass spectrometry (MS)-based proteomics explores in-depth proteome, however, the analysis of cell subsets, which might be minor population, is hampered due to a multistep procedure for sample preparation and purification. Along those lines, optimization of proteomics workflow toward limited number of cells is an asset. Here, we report our workflow of microproteomics, proteomics for a limited number of cells: a microfluidic-chip based cell sorter was applied for a damage-free cell sorting, and each 100 or 1000 THP-1 cells were collected in hydrophilic or albumin-coated microtubes. Following in-solution enzymatic digestions, peptide mixtures were purified and applied to nanoLC-MS (Orbitrap Elite) analysis with 3 hr gradient. Our method enabled proteome mapping from 1000-sorted cells, with a similar in-depth map from 1 μg of cell lysate. This approach would be useful for cell subsets and limited materials such as circulating tumor cells, inflammatory cells in cancer tissues and clinical materials. |
| Mode | partial |
| PubMed ID(s) | 28556466 |
| PubMed Info | Kasuga K, Katoh Y, Nagase K, Igarashi K Microproteomics with microfluidic-based cell sorting: Application to 1000 and 100 immune cells. Proteomics. 2017 Jul;17(13-14) |
| Principal investigator | Kie Kasuga |
| Created date | 2016-07-22 |
| Announcement date | 2017-07-18 |
| Note | - |
| Contact | Kie Kasuga Kanazawa University |
| Revision | 0 |
| Files | Files |