Fig. 6. Phase partition of n-chagasin following Triton X-114 extraction. Cells representing the three developmental stages of T. cruzi (epimastigotes, amastigotes and trypomastigotes) were lysed in 100 mM Tris-HCl buffer, pH 8.0, 150 mM NaCl, 10 mM EDTA, 0.2% of pre-condensed Triton X-114, 0.1% (w/v) NaN₃, 5 mM benzamidinium chloride and incubated with GPI-PLC (+) or not (-), for 30 minutes at 30°C. After subjecting these lysates to phase partition in Triton X-114, the samples were run on 15% SDS-PAGE and analyzed by western blotting, using rabchagasin serum. A, aqueous phase; D, detergent phase.