Table 2.
Purification of hPCNA and hCypA. Fractionation was performed on cell pellets obtained from 1 l of E. coli culture.
| Fraction | Total protein (mg)a | Purity (%)b |
|---|---|---|
| hCypA – Traditional | ||
| Supernatant | 498 | 6 |
| Pooled SP HP fractions | 5.8 | 88 |
| Pooled S-200 fractions | 5.2 | ⩾93 |
| hCypA – Automated | ||
| Pooled S-200 fractions. | 4.9 | ⩾93 |
| hPCNA – Traditional | ||
| Supernatant | 630 | 11 |
| Pooled Q HP fractions | 71.4 | 32 |
| Pooled Heparin HP fractions | 12.7 | 89 |
| Pooled S-200 fractions | 2.5 | ⩾93 |
| hPCNA – Automated | ||
| Pooled S-200 fractions | 3.6 | ⩾87 |
a
Mean values from at least 2 individual repeat runs of the corresponding protocol. Protein concentration in the supernatant and the pooled fractions after each chromatographic step was determined by BCA protein assay, apart from the final S-200 pool where protein concentration was determined by A280 measurements.
b
Determined by densitometry of appropriate lanes on reducing SDS–polyacrylamide gels (4–20% gradient).