Rat IL1 Receptor 1 protein, His tag

IL1R1 (Interleukin 1 Receptor Type I), also known as CD121a, is an important mediator involved in many cytokine induced immune and inflammatory responses. It belongs to the interleukin-1 receptor family, and is a receptor for interleukin 1 alpha (IL1A), interleukin 1 beta (IL1B), and interleukin 1 receptor antagonist (IL1RA). Besides, mouse IL1B shares 87.0% AA sequence identity with rat IL1B, suggesting the exist of cross-species activity. Thus, a binding ELISA assay was constructed to detect the association of recombinant rat IL1R1 with recombinant mouse IL1B. Briefly, IL1R1 were diluted serially in PBS with 0.1% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to IL1B-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-IL1R1 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 37ºC. Finally, add 50 μl stop solution to the wells and read at 450nm immediately. The binding activity of IL1R1 with IL1B was in a dose dependent manner.