Table 6.
Plate with contents used for the automated phosphopeptide enrichment on KingFisher Duo or Presto models
| KingFisher 96 deep-well plate order | |||
|---|---|---|---|
| Plate row number and name | μL/ well | Buffer | Time per step (min) |
| H: Tip plate | |||
| G: MagReSyn Ti-IMAC HP or Zr-IMAC HP beads | |||
| (1:2 ratio bead to peptide) | Varies based on peptide amount | 20% EtOH | |
| F: Microsphere equilibration | 500 | Phosphopeptide Bind buffer | 5 |
| E: Peptides (or TMT-labeled peptides) | 200 | Phosphopeptide Bind buffer | 20 |
| D: Wash | 500 | Phosphopeptide Bind buffer | 2 |
| C: Wash | 500 | Phosphopeptide Wash Buffer 1 | 2 |
| B: Wash | 500 | Phosphopeptide Wash Buffer 2 | 2 |
| A: Phosphopeptide elution | 200 | Phosphopeptide Elution Buffer | 10 |