Milliplex map mouse th17 magnetic bead panel kit

Th17 cells were sorted as described and plated at 10⁴ cells per well. At 24 h cytokines were measured in the supernatant by MILLIPLEX^® MAP mouse Th17 Magnetic Bead Panel kit (Merck). For co-culture experiments Th17 cells were plated at 10⁴ cells per well with dendritic cells at 3:1 ratio, together with 2.5 μM SFB peptide or 1 μg/mL EspA.

Serum samples were collected as described (Al-Homsi et al., 2017a (link), b (link)). Serum cytokines were measured using the MILLIPLEX multifactor detection technique to simultaneously analyze four inflammatory cytokines. MILLIPLEX MAP Mouse Th17 Magnetic Bead Panel kit (Merck Millipore, Billerica, MA, United States) was used for sample preparations as described (Al-Homsi et al., 2017a (link), b (link)). The expression of inflammatory factors interleukin 6 (IL-6), IL-12 (P70), IL-23, and tumor necrosis factor α (TNF-α) was detected with a BIO-RAD liquid chip device (Bio-Plex 200 system) (Hercules, CA, United States).

Intestinal tissue pieces (0.5–1 cm length) were cultured for 24ch in complete IMDM medium. Cytokines were measured in the supernatant by MILLIPLEX^® MAP mouse Th17 Magnetic Bead Panel kit (Merck) and concentrations were normalized to the weight of the explants.