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HT Colorimetric PARG Assay Kit

Poly(ADP-ribose) glycohydrolase (PARG) degrades poly(ADP-ribose) (PAR) polymers synthesized by poly(ADP-ribose) polymerase (PARP-1). When activated by DNA strand breaks, PARP-1 uses NAD as a substrate to form ADP-ribose polymers on itself and on specific acceptor proteins such as histones, DNA polymerases, DNA ligases, p53, and Fos. These polymers are in turn rapidly degraded by PARG, a ubiquitously expressed exo- and endoglycohydrolase. Excessive activation of PARP-1 leads to NAD depletion and cell death during ischemia and other conditions that generate extensive DNA damage. PARG may maintain the active state of PARP-1 by continuously removing inhibitory ADP-ribose residues from PARP-1. The regulation of PARG activity may therefore, influence PARP-mediated cell death. First, PARG inhibition could slow the turnover of PAR and thereby limit NAD depletion. Second, PARG inhibition could prevent the removal of PAR from PARP-1. Because PARP-1 is inhibited by extensive poly-(ADP-ribosyl)ation, PARG inhibitors could thereby indirectly inhibit PARP-1 activity. Prior work has shown that the PARG inhibitor gallotannin can markedly reduce death of astrocytes after oxidative stress. Trevigen’s HT Colorimetric PARG Assay Kit measures the loss of biotinylated PAR from histones attached to Strip Wells in a 96 well format and is ideal for the screening of PARG inhibitors and for measuring the activity of PARG in cell extracts.

Features:
  • Colorimetric, non-radioactive format
  • High throughput 96 test size
  • Sensitivity down to 250 pg of PARG per well

  • Applications:
  • Identify inhibitors and activators of PARG activity
  • Measuring activity of PARG in cell and tissue extracts
  •  
    Components: Part # Component Size
      4668-050-01 *PARP-HSA (10 units/µl) 50 µl
      4671-096-02 *20X PARP Buffer 500 µl
      4671-096-03 *10X PARP Cocktail 300 µl
      4671-096-06 *10X Activated DNA 300 µl
      4677-096-P Histone-Coated Strip Wells (natural) 96 wells
      4680-096-01 *PARG Enzyme (1µg/ml) 300 µl
      4680-096-02 10X PARG Buffer 8 ml
      4680-096-03 *DEA, 100 mM 200 µl
      4800-30-06 Strep-HRP 30 µl
      4822-96-08 TACS-Sapphire™ 10 ml
      *Components marked with an asterisk can be stored at -20°C for one year.
     
    Storage: Components are stored at -80°C and 4°C.
     
    References: Ying W, and Swanson RA. 2000. The poly(ADP-ribose) glycohydrolase inhibitor gallotannin blocks oxidative astrocyte death. Neuroreport. 11:1385-1388.

    Oei SL, Keil C, Ziegler M. 2005. Poly(ADP-ribosylation) and genomic stability. Biochem Cell Biol. 83:263-269.
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    Ordering Information
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    Catalog #: Price:

    4683-096-K
    HT Colorimetric PARG Assay Kit,
    96 Tests
    $485.00
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