Poly ADP Ribose Glycohydrolase (PARG) is a primary hydrolase involved in the degradation of poly(ADP-ribose) (PAR). It possesses both endo-glycohydrolase and exo-glycohydrolase activity, with a preference for the latter by binding to the two most distal ADP-ribose residues within the PAR chain. These enzymatic actions produce either free PAR or mono ADP-ribose moieties, respectively. The liberated mono ADP-ribose is further metabolized into AMP and ribose 5' phosphate by enzymes such as the NUDIX family. PARG thus plays a crucial role in regulating PAR levels, which are involved in various cellular processes including DNA damage response, chromatin maintenance, and DNA replication.Besides,Activating Transcription Factor 6 (ATF6) has been identified as an interactor of PARG, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human PARG and recombinant human ATF6. Briefly, biotin-linked PARG were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μl were then transferred to ATF6-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with Streptavidin-HRP for 30min, then wells were aspirated and washed 5 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 PARG and ATF6 was shown in Figure 1, the EC50?for this effect is 0.28ug/mL.