Immunoblotting Analysis of Trichinella spiralis Proteins

Soluble worm somatic crude and ES proteins of diverse T. spiralis phases, and purified rTsPKM were separated on 10% SDS-PAGE [24 (link),42 (link)]. The proteins were transferred onto nitrocellulose (NC) membrane (Millipore, USA) in the semi-dry transfer cell (Bio-Rad, USA) [43 (link)]. The membrane was blocked with 5% skimmed milk in Tris-buffered saline containing 0.05% Tween (TBST) at 37°C for 2 h, and cut into strips. The strips were probed by various sera (1:100; anti-rTsPKM serum, infection serum and pre-immune serum) at 37°C for 2 h. After washes with TBST, the strips were incubated at 37°C for 1 h with HRP-anti-mouse IgG conjugate (1:10000; Southern Biotech, USA). After being washed again, the strips were colored using 3, 3’-diaminobenzidine tetrahydrochloride (DAB; Sigma-Aldrich) or an enhanced chemiluminescent kit (ECL, Solarbio, China) [44 (link),45 (link)].