Biological Information
Background Information:
Poly (ADP-ribose) polymerase 2 (PARP2) is an ADP-ribosyltransferase and plays a critical role in the DNA damage response, DNA metabolism, and chromatin architecture. While PARP1 is activated by multiple forms of DNA damage-mimicking oligonucleotides, PARP2 is specifically activated by 5′-phosphorylated DNA breaks. At the BER process of DNA repair, PARP1 synthesizes poly ADP-ribose (PAR) and recruits repairing machinery at the initial stage, along with PARP2 which maintains the following repair process. Pharmacological inhibition of PARP2 provides a multipronged attack on tumor physiology in genomic, cellular, and microenvironmental aspects.
Family:
DNA Repair & Mitotic Regulation
Class:
Other
Protein Name:
PARP2
Uniprot Number:
Q9UGN5
Protein Aliases:
ARTD2
Gene Name:
PARP2
Gene ID:
10038
Gene Aliases:
ARTD2
Accession Number:
NM_005484
Organism:
Human
Construct Details:
Full length, Recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Spectrophotometry
Platform:
Plate reader
Measured Response:
Absorbance
Testing Information
Procedure Summary:
Human recombinant PARP2 protein expressed in Sf9 cells is used. 100 ng histone proteins are coated on a 96-well plate. Test compound and/or vehicle is preincubated with 0.25 µg/ml enzyme for 15 minutes at 25°C in Tris-HCl buffer pH 8.0. The reaction is initiated by addition of 25 µM spiked biotinylated NAD+ plus NAD+ for another 60 minutes reaction time. Streptavidin-HRP is added and followed by addition of the TMB substrate to produce color that can be measured at 450 nm on a spectrophotometer. Compounds are screened at 10 µM.
Substrate:
25 µM NAD+
Incubation:
60 min at 25°C
Control Inhibitor:
Olaparib
Criteria For Significance:
≥ 50% of max stimulation or inhibition
Test Concentration / Dose:
Suggested concentration for initial testing is 10 uM
Test Sample Requirements:
Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).
Minimum Order Quantity:
2
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Clinical Relevance
Therapeutic Area:
Oncology/Immuno-Oncology
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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