TABLE 1.
iTRAQ label | Protein quantity to be used in pooling (μg) | Approximate volume (μL) of 100 μg digested proteins after Speed Vac | Ethanol to add to labeling reaction (μL) | Approximate volume of iTRAQ labeling reagent (μL) | Water to add to quench labeling reaction (μL) | Volume of label and digest after quenching (μL) | Volume to match protein quantity desired in pooling (μL) |
---|---|---|---|---|---|---|---|
114 | 5 | 40 | 70 | 35 | 100 | 246 | 12.3 |
115 | 25 | 40 | 70 | 35 | 100 | 250 | 62.5 |
116 | 50 | 40 | 70 | 35 | 100 | 245 | 122.5 |
117 | 100 | 40 | 70 | 35 | 100 | Combined all labels into this vial | Combined all labels into this vial |