Active Cold Inducible RNA Binding Protein (CIRBP)

CIRBP (Cold-inducible RNA-binding protein) is considered to play a protective role in the genotoxic stress response by stabilizing transcripts of genes involved in cell survival and act as a translational activator. Besides, ATXN1 (Ataxin-1) has been proven as an interactor of CIRBP. Thus a binding ELISA assay was conducted to detect the interaction of recombinant human CIRBP and recombinant human ATXN1. Briefly, CIRBP were diluted serially in PBS, with 0.01%BSA (pH 7.4). Duplicate samples of 100uL were then transferred to ATXN1-coated microtiter wells and incubated for 2h at 37°C. Wells were washed with PBST and incubated for 1h with anti-CIRBP 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 CIRBP and ATXN1 was shown in Figure 1, and this effect was in a dose dependent manner.