Figure. The binding activity of IFNa with IFNa/bR1.
Interferon Alpha (IFNa) belongs to a large subgroup of interferon proteins that help regulate the activity of the immune system. The IFNa proteins are produced by leukocytes. They are mainly involved in innate immune response against viral infection. It is also made synthetically as medication in hairy cell leukemia. Besides, Interferon Alpha/Beta Receptor 1 (IFNa/bR1) has been identified as an interactor of IFNa, thus a binding ELISA assay was conducted to detect the interaction of recombinant rat IFNa and recombinant rat IFNa/bR1. Briefly, IFNa were diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to IFNa/bR1-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-IFNa 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 50µL stop solution to the wells and read at 450nm immediately. The binding activity of IFNa and IFNa/bR1 was shown in Figure 1, and this effect was in a dose dependent manner.