Materials and Reagents

1. Paraformaldehyde (PAF) (Sigma-Aldrich, catalog number: 158127 )
   
2. PBS (Sigma-Aldrich, catalog number: P4417 )
   
3. BSA (Sigma-Aldrich, catalog number: A3803 )
   
4. poly-D-Lysine–coated slides (culture slides) (BD Biosciences, Falcon^®, catalog number: 354632 )
   
5. Mouse anti-HisG monoclonal antibody (Life Technologies, catalog number: R940-25 )
   
6. Alexa Fluor 594 goat anti-mouse IgG (Life Technologies, catalog number: A 110005 )
   
7. Dako fluorescent mounting medium (Dako, catalog number: S3023 )
   
8. Primary sporocysts of S. mansoni (Brazilian strain) used for immunocytochemical experiments were obtained by transferring miracidia to Chernin’s balanced salt solution (CBSS) and maintaining at 26 °C under normoxic conditions for 24 h (Yoshino and Laursen, 1995). Then, 100 primary sporocysts were incubated for 1 h with 30 nM of recombinant Biomphalysin protein (Galinier et al., 2013). As negative control, 100 sporocysts were used without having been treated with recombinant biomphalysin.
   
9. N terminal His(6)-tagged biomphalysin was expressed in vitro using the Rapid Translation System (RTS 500 Wheat Germ CECF Kit) (5 PRIME, catalog number. 2402500 )
   
10. 4% PAF (see Recipes)
    
11. PBS/3% BSA (see Recipes)
    
12. PBS/1% BSA (see Recipes)
    
13. Anti- HisG antibody 1:500 (see Recipes)
    
14. Anti-mouse IgG 1:1,000 (see Recipes)
    

Equipment

1. Coverslip 24 x 60 mm (VWR International, catalog number: 631-1575 )
   
2. BD BioCoat^TM Poly-D-Lysine 8-well CultureSlides (Becton, Dickinson and Company, catalog number: 354632)
   
3. Eppendorf centrifuge (Eppendorf , model: 5810R )
   
4. Swinging agitator (Fisher scientific, model: 10758995 )
   
5. Fluorescence confocal laser-scanning microscope (ZEISS, model: LSM 700 )
   

Procedure