Ready set go elisa sets

Splenic CD4⁺ T cells were isolated from Ova-TCR transgenic DO11.10 mice using the CD4⁺ T Cell Isolation Kit (Miltenyi Biotec, Bergisch Gladbach, Germany). BALB/c mDCs (3.2 × 10⁵ cells/mL) were cultured alone or in combination with DO11.10 CD4⁺ T cells (8.0 × 10⁵ cells/mL, >95% purity) and stimulated with equimolar amounts of Ova, MPLA, MPLA mixed with Ova (MPLA + Ova), or MPLA : Ova (fusion protein) for 72 h. Subsequently, concentrations of IL-1β, IL-2, IL-5, IL-6, IL-9, IL-10, IL-12p70, IL-13, IL-17A, TNF-α, and IFN-γ in the supernatants were measured by BD OptEIA ELISA (BD Biosciences, Heidelberg, Germany) or Ready-SET-Go! ELISA Sets (eBiosciences, Frankfurt, Germany).

Splenic CD4⁺ T cells were isolated from OVA-TCR transgenic DO11.10 mice using the CD4 T Cell Isolation Kit (Miltenyi Biotec, Bergisch Gladbach, Germany). BALB/c-derived BMmDCs (3.2 × 10⁵ cells/mL) were either cultured alone or together with DO11.10 CD4⁺ T cells (8.0 × 10⁵ cells/mL, >95% purity) and stimulated with equimolar amounts of Pam₂CysK₄, CL264, Pam₂CysK₄ plus CL264, CL401, CL413, or CL531 in the presence or absence of 20 μg/mL rOVA for 72 h. Subsequently, concentrations of IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-13, IL-17A, TNF-α, and IFN-γ in the supernatants were measured by either BD OptEIA™ ELISA (BD Biosciences) or Ready-SET-Go! ELISA Sets (eBiosciences).