Fig 1: Importance of Dp1 and antigen fusion without DC activation(A) Mice were immunized with 1 µg PspA alone, a mixture of 1 µg PspA and 1 µg free Dp1×3, or 1 µg PspA plus 250 µg alum, and the level of PspA-specific IgG in the plasma samples was evaluated at 7 days after the second immunization using ELISA.(B) Mice were immunized with 10 µg OVA alone, 10 µg OVA plus 1 µg PspA-Dp1×3, or 10 µg OVA plus 250 µg alum, and the level of OVA-specific IgG in the plasma samples was evaluated 7 days after the second immunization using ELISA.(C) Mice were immunized with 1.1 nmol (equivalent to 5 µg) OTII-Dp1×3, 1.1 nmol (equivalent to 5 µg) OTII-Dp1×3 plus 4 nmol (equivalent to 100 µg) free Dp1×3, or 1.1 nmol (equivalent to 5 µg) OTII-Dp1×3 plus 4 nmol (equivalent to 100 µg) free Dp5×3, and the level of OVA-specific IgG in the plasma samples was evaluated at 7 days after the second immunization using ELISA. (A–C) We used (A and B) 800- (?), 4,000- (?), and 20,000- (?) fold–diluted, and (C) 160- (?), 800- (?), and 4,000- (?) fold–diluted plasma samples.(D) BMDCs were treated with 11.3 nM (equivalent to 50.9 µg/mL) OTII-Dp1×3, 11.3 nM (equivalent to 46.3 µg/mL) OTII-Dp2×3, 11.3 nM (equivalent to 20 µg/mL) OTII, or 11.3 nM (equivalent to 20 µg/mL) OTII plus 10 µg/mL CpG ODN for 24 h, and the levels of IL-6 and IL-12 p40 in the supernatants were determined using ELISA. (A and B) n = 4; (C) n = 5; (D) n = 4. Data are means ± SD. *p< 0.05, ***p< 0.001, ****p< 0.0001 as indicated by Tukey’s test. N.S.: not significant. Significant differences were analyzed only in the (A and B) 800-fold-diluted and (C) 160-fold-diluted plasma samples.
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