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1. Spin out any remaining debris in ascites 15 mins., 3K in SS34.
2. Set up beaker in ice bath on stirring plate.
3. Measure volume of ascites (+ 0.5 mLs).
4. Pour into beaker and start stir bar going rapidly.
5. Measure out equal volume of 50% NH4SO4.
6. Add slowly to ascites fluid.
7. Stir on ice 1 hr. - precipitate should form.
8. Leave at 4°C (no stirring) overnight.
9. Spin 30 minutes in Sorvall centrifuge (3000g), in HB-4.
10. Discard supernatant.
11. Resuspend pellet in minimum volume HBSS (i.e., 5 mL for 15-20 mL ascites).
12. Dialyze against PBS at 4°C for 24 hrs. 2-3 changes.
13. Filter through 0.45 mm filter.
14. Can use this way as crude ascites or purify further.
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