Mouse Annexin II protein, His and TrxA tag

Annexin A2 (ANXA2) also known as annexin II is a member of the annexin family. Members of this calcium-dependent phospholipid-binding protein family play a role in the regulation of cellular growth and in signal transduction pathways. This protein functions as an autocrine factor which heightens osteoclast formation and bone resorption. It is a calcium-dependent phospholipid-binding protein whose function is to help organize exocytosis of intracellular proteins to the extracellular domain. Besides, S100 Calcium Binding Protein A10 (S100A10) has been identified as an interactor of ANXA2, thus a binding ELISA assay was conducted to detect the interaction of recombinant mouse ANXA2 and recombinant mouse S100A10. Briefly, ANXA2 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to S100A10-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-ANXA2 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 ANXA2 and S100A10 was in a dose dependent manner.