武汉云克隆科技股份有限公司

Serum Amyloid A (SAA) is a class of 104 amino acid conservative acute-phase proteins, which is essential in immune-mediated inflammatory processes, like IBD. SAA has dual roles in the inflammatory process, which it can stimulates the pro-inflammatory cytokine expression and promotes the pathogenic differentiation of TH17 cells. In addition, SAA can remove toxic lipids produced during inflammatory responses and membrane debris from dead cells, redirect HDL, and recycle cholesterol for tissue repair. Besides, C Reactive Protein (CRP) has been identified as an interactor of SAA, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human SAA and recombinant human CRP. Briefly, SAA were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 ul were then transferred to CRP-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-SAA 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 SAA and recombinant human CRP was shown in Figure 1, the EC50 for this effect is 5.3 ug/mL.