Mouse ifn β elisa kit

DCs were seeded in 12-well plates with 2 × 10⁵ cells/well and incubated for 24 h. The following treatments were performed on these cells: (1) un-treated; (2) adding CpG at a concentration of 0.5 μg/mL; (3) adding PIC at a concentration of 4.67 μg/mL. Twenty four hours later, the supernatants were collected for the quantitative analysis of IFN-β using a mouse IFN-β Elisa kit (Biolegend, Cat # 439407), and the cells were collected and stained with antibodies (or dyes): anti-CD11c PerCP-Cy5.5, anti-CD80 PE, anti-CD86 BV605, anti-CD317 Alexa 700, Live/Dead Ghost Red 780. To study the cellular uptake of FITC-Ova, the following treatments were performed on the DCs: (1) un-treated; (2) adding FITC-Ova at a concentration of 1.67 μg/mL. (3) adding the mixture of FITC-Ova and PIC (1.67 μg/mL FITC-Ova and 4.67 μg/mL PIC). The FITC-Ova and PIC were mixed 20 min before adding to the cells. Twenty four hours later, the cells were collected and stained with anti-CD11c PerCP-Cy5.5 and Live/Dead Ghost Red 780. The UltraComp Beads eBeads (Invitrogen) were used for compensation for flow cytometry. All samples were incubated with CD16/CD32 (Fc block) for 5 min at room temperature before staining. Flow cytometry was performed on an Attune Cytometer (ThermoFisher).

IFNβ secretion by mixed glial cells upon stimulation with LPS/U0126 was measured using a commercially available mouse IFNβ ELISA kit (Biolegend). ELISA was performed according to the manufacturer’s instructions.