Poly (ADP-ribose) Polymerase 1 (PARP1) Human Enzymatic LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Poly (ADP-ribose) polymerase 1 (PARP1), the first member of the PARP family, post-translationally ADP-ribosylates itself, other proteins, and DNA. PARP1 uses NAD+ as a substrate to synthesize poly ADP-ribose (PAR) and transfers PAR to target proteins. PARP1 has diverse roles in different cellular processes, such as differentiation, proliferation, tumor transformation, and DNA repair. PARP1 is overexpressed in the progression of endometrial cancer, BRCA-mutated ovarian cancer, and other cancers. The inhibitor of the catalytic sites of PARP1 is therefore being exploited clinically to treat various cancers, including DNA repair-deficient ovarian, breast, and prostate cancers.
Family:
DNA Repair & Mitotic Regulation
Class:
Other
Protein Name:
PARP1
Uniprot Number:
P09874
Protein Aliases:
ARTD1
Gene Name:
PARP1
Gene ID:
142
Gene Aliases:
ARTD1; PARS
Accession Number:
NM_001618
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 PARP1 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 12.5 µM spiked biotinylated NAD+ plus NAD+ for another 60 minutes of 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:
12.5 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
RELATED SOLUTIONS
PARP Screen LeadHunter Panel - TW
Item: PP317
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