Tumor protein p53 (TP53), also known as p53, cellular tumor antigen p53 has many mechanisms of anticancer function and plays a role in apoptosis, genomic stability, and inhibition of angiogenesis. TP53 act as a tumor suppressor in many tumor types; induces growth arrest or apoptosis depending on the physiological circumstances and cell type. It also Involved in cell cycle regulation as a trans-activator that acts to negatively regulate cell division by controlling a set of genes required for this process. Besides, CREB Binding Protein (CREBBP) has been identified as an interactor of TP53, thus a binding ELISA assay was conducted to detect the interaction of recombinant rat TP53 and recombinant rat CREBBP. Briefly, TP53 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μL were then transferred to CREBBP-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-TP53 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 50μL stop solution to the wells and read at 450nm immediately. The binding activity of TP53 and CREBBP was shown in Figure 1, and this effect was in a dose dependent manner.
Figure. The binding activity of TP53 with CREBBP.