Ribonuclease P (RNASEP) is a type of ribonuclease which cleaves RNA. RNase P is unique from other RNases in that it is a ribozyme – a ribonucleic acid that acts as a catalyst in the same way that a protein based enzyme would. Its function is to cleave off an extra, or precursor, sequence of RNA on tRNA molecules. Besides, Methyl CpG Binding Protein 2 (MECP2) has been identified as an interactor of RNASEP, thus a binding ELISA assay was conducted to detect the interaction of recombinant human RNASEP and recombinant human MECP2. Briefly, RNASEP were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μL were then transferred to MECP2-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-RNASEP 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 RNASEP and MECP2 was shown in Figure 1, and this effect was in a dose dependent manner.
Figure. The binding activity of RNASEP with MECP2.