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. 2022 Nov 4;12(21):3040. doi: 10.3390/ani12213040

Figure 2.

Figure 2

Checkerboard titration plate layout. The capture antibody (cAb) is titrated along rows and the detection antibody (dAb) along columns, as indicated. Rows D and H and columns 6 and 12 are blank (BL) and do not contain antibodies. To each of the 4 quarters of the plate one concentration of cytokine is added (10,000, 1000, 100, or 0 pg/mL). This setup allows to test the combination of multiple cAb and dAb concentrations with cytokines in one go, using one ELISA plate. Starting values (1:1) for antibodies for IL-2, IL-6 and IL-10 was 2.5 µg/mL cAb and 0.2 µg/mL dAb, for IL-12p40 was 2 µg/mL cAb and 0.02 µg/mL dAb, and for IFN-γ was 4 µg/mL cAb and 0.5 µg/mL dAb.