Table 3.27.1.
Mitochondrial and ER Yields and Purity from Differentially Lysed HeLa Cells
| Average
| ||||||||
|---|---|---|---|---|---|---|---|---|
| Lysis method | Yield samples | Initial cell pellet (g) | Total protein fraction (mg) | Average Mitochondria yield (μg) | ER yield (μg) | Yield | MAM detected in | Purity |
| Sonication | ||||||||
| Low (3 × 5 sec) | 1, 2 | 0.153 | 9.87 | 415.86 | 842.6 | High | ER + mitochondria | Good |
| Medium (3 × 10 sec) | 1, 2 | 0.148 | 9.584 | 350.34 | 824.4 | High | ER | Good |
| High (3 × 15 sec) | 1, 2 | 0.146 | 8.778 | 163.14 | 820.5 | Medium | ER | Good |
| Freeze/thaw | 1, 2 | 7.634 | ND | 195.2 | Medium | ER + mitochondria | Good | |
| Homogenization | 1, 2 | 0.146 | 1.484 | NDa | NDa | Low | MAM/mitochondria | Excellent |
| 3, 4 | 0.147 | 1.392 | NDa | NDa | Requires large scale-up | Mitochondria | ||
a
ND, not determined as extract protein concentrations were below detection level of the BCA assay.