PARG Fluorogenic Assay Kit
The PARG Fluorogenic Assay Kit is a high-throughput, homogeneous 96 or 384 well assay designed to measure the hydrolase activity of Poly (ADP-ribose) glycohydrolase (PARG) for screening and profiling applications, using a simple and straightforward fluorogenic assay. The PARG Fluorogenic Assay Kit contains enough purified recombinant PARG enzyme, substrate, and assay buffer for 100 or 400 enzyme reactions. The kit also includes the PARG inhibitor PDD00017273 as a PARG activity control.
Figure 1: Illustration of the assay principle.
PARG is incubated with a fluorogenic ADP-ribose substrate in which the fluorophore is quenched by the presence of the ribose. PARG-mediated hydrolysis of the substrate between the ribose and the fluorochrome releases fluorescence that can be detected at λ=502 nm (excitation at λ=385 nm). Fluorescence intensity is directly proportional to PARG hydrolase activity.
Need us to run inhibitor screens or profile your compounds against PARG? Check out our PARG Screening Services or DNA Replication and Repair Screening Services.
- Fluorescence plate reader capable of excitation at 385 nm and detection at 502 nm
- Adjustable micropipettor and sterile tips
- Rotating or rocker platform
96 Reactions
| Catalog # | Name | Amount | Storage |
| 101726 | PARG, Full Length, His-Tag* | 0.5 µg | -80°C |
| 82138 | Fluorogenic PARG Substrate (1 mM) | 25 µl | -20°C |
| Stock PARG Hydrolase Buffer | 1.2 ml | -20°C | |
| 0.5 M DTT | 200 µl | -20°C | |
| PDD00017273 (10 mM) | 20 µl | -20°C | |
| 79685 | 96-well microplate, black | 1 plate | Room Temp |
*The concentration of the protein is lot-specific and will be indicated on the tube.
384 Reactions
| Catalog # | Name | Amount | Storage |
| 101726 | PARG, Full Length, His-Tag* | 1 µg | -80°C |
| 82138 | Fluorogenic PARG Substrate (1 mM) | 40 µl | -20°C |
| Stock PARG Hydrolase Buffer | 3 ml | -20°C | |
| 0.5 M DTT | 200 µl | -20°C | |
| PDD00017273 (10 mM) | 20 µl | -20°C | |
| 79961 | 384-well microplate, black | 1 plate | Room Temp |
*The concentration of the protein is lot-specific and will be indicated on the tube.
Poly (ADP-ribose) glycohydrolase (PARG) is a catabolic enzyme involved in the degradation of PARylated chains, releasing ADP-ribose and oligo (ADP-ribose) chains. PAR homeostasis is regulated by the family of PAR polymerases (PARPs) and PARG in response to cellular stress conditions such as DNA damage response (DDR). PARG activity is linked to cellular responses in inflammation, ischemia, stroke, and cancer. PARG is overexpressed in breast cancer and associated with tumor growth and survival. Decrease in PARG activity can potentiate the effect of current cancer therapies, such as chemotherapy and radiation, making PARG inhibition with selective inhibitors a promising approach in cancer and immunotherapy.
Marques, M. et al. 2019 Oncogene 38 (12): 2177-2191.
James, D. I. et al., 2016 ACS Chem Biol 11 (11): 3179-3190.
Drown, B. S. et al., 2018 Cell Chem Bio 25 (12): 1562-1570.
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