PARG Fluorogenic Assay Kit

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Catalog #
78858
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Description

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.

Synonyms
Poly(ADP-ribose) glycohydrolase, PARG99
Product Info
Storage and Usage
Citations
Assay Kit Format
Fluorogenic
Materials Required But Not Supplied
  • Fluorescence plate reader capable of excitation at 385 nm and detection at 502 nm
  • Adjustable micropipettor and sterile tips
  • Rotating or rocker platform
Format

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.

UniProt #
Q86W56
Background

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.

References

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.