Mouse il 17a elisa max kit

MNCs isolated from spleens of mice infected with TMEV were cultured with 2
× 10⁶ cells/well in 6-well plates (Corning Inc., Corning, NY) and
stimulated with 5 μg/ml of concanavalin A (ConA) or 1 MOI of TMEV for 48 hours.
After 2 days the supernatants were collected and analyzed by enzyme-linked immunosorbent
assay (ELISA) (42 (link)). The levels of IL-4, IL-6,
IL-10, and IFN-γ concentrations were detected with BD OptEIA kits (BD Biosciences,
San Diego, CA) and IL-17A was detected with a Mouse IL-17A ELISA MAX^™kit (Biolegend, Inc., San Diego, CA) according to the manufacturer’s instructions
(45 (link)). ELISAs were performed in duplicate in
96-well plates.

A volume of 2 ml of 4 × 10⁶ spleen MNCs in RPMI 1640 supplemented with 1% glutamine, 1% antibiotics, 50 μM 2-mercaptoethanol, and 10% FBS was added to each well of 6-well plates. This was incubated with 2 ml of media or a solution containing 10 μg/ml ConA or 4 × 10⁶ PFUs of TMEV MOI of 1 for 2 days. After 2 days the supernatants were collected and analyzed by ELISA (58 (link)). IFN-γ, IL-4, and IL-10 concentrations were detected with BD OptEIA kits (BD Biosciences) and IL-17 was detected with a Mouse IL-17A ELISA MAX^™ kit (BioLegend).

MNCs isolated from inguinal lymph nodes of mice were cultured at 8 × 10⁶ cells/well in 6-well plates (Corning Inc., Corning, NY) and stimulated with 4 ml of 5 μg/ml of concanavalin A (ConA) or 50 μg/ml of MOG_35-55 for 48 hr. Culture supernatants were harvested and stored at −80 °C until examined. The levels of IL-4, IL-6, IL-10, and IFN-γ concentrations were detected with BD OptEIA kits (BD Biosciences, San Diego, CA) and IL-17A was detected with a Mouse IL-17A ELISA MAX^™ kit (Biolegend, Inc., San Diego, CA) according to the manufacturer’s instructions (Tsunoda et al., 2005 (link)). ELISAs were performed in duplicate in 96-well plates.