S100A6 is a calcium-binding protein, which functions as calcium sensor and modulator, contributing to cellular calcium signaling. It is reported that S100B and S100A6 differentially modulate cell survival by Interacting with distinct RAGE (Receptor for Advanced Glycation End Products). Thus a binding ELISA assay was conducted to detect the interaction of S100A6 and RAGE. Briefly, recombinant human S100A6 were diluted serially in PBS, with 0.01%BSA (pH 7.4). Duplicate samples of 100uL were then transferred to RAGE-coated microtiter wells and incubated for 2h at 37°C. Wells were washed with PBST and incubated for 1h with anti-S100A6 mAb, 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°C. Finally, add 50μL stop solution to the wells and read at 450nm immediately. The binding activity of of S100A6 and RAGE was shown in Figure 1, and this effect was in a dose dependent manner.