Goat anti mouse igg fitc conjugate

Whole worms of different T. spiralis stages (ML, IIL, AW, and NBL) were fixed with 4% formaldehyde for 30 min and re-fixed with cold acetone for 20 min. The fixed worms were embedded in paraffin and cut into a 2-μm-thick cross-section with a microtome. The expression and tissue location of native TsGS in various worm stages were investigated by indirect immunofluorescent assay (IIFA) (Liu et al., 2017 (link); Cui et al., 2019 (link)). Whole parasites and their cross-sections were blocked with 5% normal goat serum for 2 h and then probed at 37°C for 2 h with 1:10 diluted diverse sera (anti-rTsGS serum, infection serum, and normal serum). After washes with PBS, the worms were stained using goat anti-mouse IgG-FITC conjugate (1:100; Santa Cruz, United States). Following washes again, whole worms and cross-sections were examined under a fluorescence microscope (Olympus, Tokyo, Japan) (Lei et al., 2020 (link); Yue et al., 2020 (link)).

All the slides containing cells were fixed in 4% paraformaldehyde for 15 min, permeated in 0.1% Triton X-100 for 5 min, and then blocked in 5% goat serum for 1 h. HASMCs were stained or costained with rabbit anti-TRAF6 (Cat#ab62488, Abcam; 1:50) and mouse anti-NF-κB (Cat#sc8008, Santa Cruz; 1:50) overnight at 4°C. The cells were incubated with secondary goat anti-rabbit IgG-PE conjugate (Cat#sc3739, Santa Cruz; 1:100) or goat anti-mouse IgG-FITC conjugate (Cat#sc2010, Santa Cruz; 1:100). The slides were washed 3 times in between each step, and lastly, incubated with anti-fade reagent with DAPI (Cat#8961, CST, USA). The cells were observed under a confocal laser scanning microscopy (LSM710, ZEISS, Germany). We took 4 images for every slide randomly, and repeated all the experiments for at least 3 times.