Spectrometer nd 1000 uv vis

H. diminuta adult tapeworms and cysticercoids were suspended in lysis buffer (8 M Urea, 4% CHAPS, 40 mM Tris base) for homogenisation by sonication on ice until the suspension became clear. The homogenate was centrifuged at 54,782 × g at 4°C for 25 min to collect the supernatant containing the solubilised proteins, which were either used directly for the present study or stored at −80°C until use. The amount of total protein was measured using a Spectrometer ND-1000 UV/Vis (NanoDrop Technologies, USA). Samples used for biological replicates for in-gel and in-solution digestion were taken from independent experiments.

Hymenolepis diminuta adult worms in whole (size between 40 and 60 cm in length) were suspended in lysis buffer, containing 8 M Urea, 4% CHAPS and 40 mM Tris-base supplemented with protease inhibitor cocktail (Roche, Germany) and homogenized by sonication on ice until the suspension became clear. The homogenate was centrifuged at 15,000 × g at 4°C for 25 min to collect the supernatant containing soluble proteins, which were either used directly or stored at −80°C until use. The protein concentration was measured using a Spectrometer ND-1000 UV/Vis (NanoDrop Technologies, United States). Three biological replicates (three adult worms at the same age collected from different animals) taken from independent experiments were used in the present study.

After thawing, H. diminuta cysticercoids were again extensively washed three times in PBS (100 mM) and then mixed with lysis buffer (8 M Urea, 4% CHAPS, 40 mM Tris-base, supplemented with protease inhibitor cocktail; Roche, Berlin, Germany) to solubilize protein components. Then the protein mixture was homogenized in a glass Potter-homogenizer and disintegrated by sonication. The lysis solution was clarified by centrifugation at 14,000× rpm for 15 min in an Eppendorf microcentrifuge. Concentration of proteins was measured using the Spectrometer ND-1000 UV/Vis (NanoDrop Technologies, Wilmington, USA). Proteins were kept at -80 °C for further analysis.