Active Interleukin 12A (IL12A)

human IL12A is a subunit of interleukin 12, a cytokine that acts on T and natural killer cells, and has a broad array of biological activities. Interleukin 12 is a disulfide-linked heterodimer composed of the 40 kD cytokine receptor like subunit encoded by this gene, and a 35 kD subunit encoded by IL12A. This cytokine is expressed by activated macrophages that serve as an essential inducer of Th1 cells development. Interleukin 12A can combind with Epstein Barr Virus Induced Protein 3 (EBI3). Thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human IL12A and recombinant human EBI3. Briefly, IL12A were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 µl were then transferred to EBI3-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-IL12A pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450 nm immediately. The binding activity of recombinant human IL12A and recombinant human EBI3 was shown in Figure 1, the EC50 for this effect is 38.3 ng/mL.