Table 1.
Purification of GPI-anchored protein releasing activity from male germ cells
| Column | Volume (ml) | Total protein (mg) | Total activity (AU)* | Specific activity (U/mg protein) | Fold purification |
|---|---|---|---|---|---|
| S-100 sup. | 393 | 19,100 | 336,160 | 18 | 1.0 |
| DEAE-cellulose | 8 | 228 | 14,410 | 63 | 3.5 |
| Phenyl-sepharose | 8 | 212 | 17,596 | 83 | 4.6 |
| Con-A-sepharose | 1 | 6 | 6,100 | 1,017 | 56.5 |
| TSK gel 3000SW | 1 | 1 | 2,500 | 2,500 | 138.9 |
The membrane-rich fraction of germ cells from mouse testis was solubilized in a buffer containing 1% Triton X-100, centrifuged and the supernatant was collected and subjected to chromatographic fractionation. The PLAP conversion assay was performed on the eluted fractions and the maximum values are used here.
All reactions were performed using PI-PLC (1.0 IU/ml), the value of which was defined as maximum reaction.