Poly ADP Ribose Glycohydrolase (PARG) is a critical enzyme responsible for the catabolism of poly(ADP-ribose) (PAR), a post-translational modification synthesized primarily by PARP enzymes in response to DNA damage. By hydrolyzing PAR chains, PARG plays an essential role in regulating DNA repair, chromatin remodeling, and cell fate decisions, such as apoptosis and necrosis. Its activity ensures the transient nature of PAR signaling, and dysregulation of PARG is implicated in cancer, neurodegeneration, and inflammation. PARG interacts with Sirtuin 1 (SIRT1), influencing its deacetylase activity and participating in the coordination of stress response and metabolic pathways.
To detect the activity of recombinant PARG , a functional ELISA assay was performed to evaluate the interaction between recombinant human PARG and recombinant rat SIRT1.
Briefly, PARG was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to SIRT1-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-PARG pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630nm immediately. The binding activity of recombinant human PARG and recombinant rat SIRT1 was shown in Figure 1, the EC50 for this effect is 0.019µg/mL..