For SWA preparation, S. japonicum adult worms were suspended in cold diethyl ether and homogenized on ice using a homogenizer (VirTis Co., Inc., Gardiner, NY). The homogenate was centrifuged at 2,000 g for 5 min at 4°C to remove lipids. The pellet was freeze-thawed several times in PBS mixed with 1 mM PMSF (Roche Diagnostics) and 2ug/ml Leupeptin (Sigma). The homogenate was centrifuged at 20,000 g for 50 min at 4°C and the supernatant was filtered through 0.22 μm filter (Millipore Corporation). Protein concentration of worm extracts were determined by BCA Protein Assay Kit (Bio-Rad) and the extracts were stored at -80°C before use [20 (link)].
To prepare SEA, S. japonicum eggs were suspended in PBS containing 1 mM PMSF (Roche Diagnostics) and 2 μg/ml Leupeptin (Sigma) and homogenized on ice using a homogenizer (VirTis Co.). The suspension was freeze-thawed several times and centrifuged at 20,000 g for 50 min at 4°C. The supernatant was filtered through 0.22 μm filter (Millipore Corporation). Protein concentration of egg extracts were determined by BCA Protein Assay Kit (Bio-Rad) and the extracts were stored at -80°C before use [21 (link)].
To prepare SEA, S. japonicum eggs were suspended in PBS containing 1 mM PMSF (Roche Diagnostics) and 2 μg/ml Leupeptin (Sigma) and homogenized on ice using a homogenizer (VirTis Co.). The suspension was freeze-thawed several times and centrifuged at 20,000 g for 50 min at 4°C. The supernatant was filtered through 0.22 μm filter (Millipore Corporation). Protein concentration of egg extracts were determined by BCA Protein Assay Kit (Bio-Rad) and the extracts were stored at -80°C before use [21 (link)].
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