Mouse CD47 protein, His tag

Integrin Associated Protein (IAP) also known as Cluster of Differentiation 47(CD47) is a transmembrane protein. CD47 belongs to the immunoglobulin superfamily and partners with membrane integrins and also binds the ligands thrombospondin-1 (TSP-1) and signal-regulatory protein alpha (SIRPα). CD47 is involved in a range of cellular processes, including apoptosis, proliferation, adhesion, and migration. Furthermore, it plays a key role in immune and angiogenic responses. CD47 is ubiquitously expressed in human cells and has been found to be overexpressed in many different tumor cells. Besides, Thrombospondin 1 (THBS1) has been identified as an interactor of IAP, thus a binding ELISA assay was conducted to detect the interaction of recombinant mouse IAP and recombinant mouse THBS1. Briefly, IAP were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to THBS1-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-IAP 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 IAP and THBS1 was in a dose dependent manner.