PARP Assays
Target-specific Assays

Poly (ADP-ribose) Polymerase (PARP) Assay Services

Poly (ADP-ribose) polymerase (PARP) is a family of proteins that transfer ADP-ribose units from NAD+ onto target nuclear proteins forming long branched Poly ADP-ribose chains. PARPs play a role in epigenetic regulation, for example, by poly ADP-ribosylation of histone substrates.

  • Enzyme activity assay
  • Low and large scale screening, as well as high-throughput screening options, are available
  • Custom assay development possible
  • PARP1 and PARP2 are produced in house and available for purchase
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List of Targets

Assay Details

  • Assay Format
  • Assay Setup
Assay Format

PARP activity transfers the ADP-Ribose units from NAD+ co-factor to histone proteins. PARP activity assay quantifies remaining NAD+ after enzymatic reaction via NAD+/NADH Glo Kit

Assay Setup

Setups: Single-dose screening in duplicates or IC50 value determination with 5 or 10 concentrations. Other screening formats are available upon request.

Controls: No inhibitor (DMSO vehicle) control and for every assay, one target-specific control compound is tested in 10-dose IC50 format.

Turnaround time: 10 business days for standard projects. Expedited scheduling and data delivery can be arranged prior to the commencement of the studies.

Report: The raw data, % enzyme activity and control compound IC50 values will be reported in Excel format for single-dose assays. For IC50 orders, raw data, IC50 values, and curve fitting will be delivered in Excel format. Assay conditions, target, and substrate information are available upon request. Requirements for this information should be noted prior to the commencement of the study.

Screening facility: This assay is performed at our screening facility in Malvern, PA, US.

Compound requirements: In brief, for a standard project, 20ul of a 10mM DMSO stock or solid material is needed. Less material is needed for large scale screening. Please refer to our FAQs for information regarding compound preparation and shipping.

Please see our FAQ page to access our assay condition form (US facility).

NanoBRET Target Engagement Assays for PARPs in Live Cells

NanoBRET Target Engagement (TE) is a biophysical method of assessing the binding between a compound and a target protein in live cells. Based on bioluminescent resonance energy transfer (BRET), NanoBRET TE can be applied to multiple classes of target proteins, including members of the poly (ADP-ribose) polymerase (PARP) family, which play critical roles in cellular processes such as transcription, cell division and DNA repair.

List of NanoBRET TE Targets

NanoBRET TE Assay Details

  • Assay Format
  • Screening Details
Assay Format

In the NanoBRET TE Assays for PARPs, the target PARP protein is expressed as a NanoLuc Luciferase fusion. A cell-permeable, fluorescent NanoBRET PARP tracer is added. The tracer reversibly binds to the PARP target-NanoLuc fusion protein. When in proximity to one another, the tracer and NanoLuc Luciferase produce a BRET signal. When a test compound binds to the PARP target proteins, it competes with the tracer, and the BRET signal is lost.

Screening Details

Setups: IC50 value determination in 10-dose duplicate assay format. Free choice for screening of the PARP targets of your choice.

Controls: We use DMSO as a negative control. In every project, testing of a standard reference compound (Olaparib) is included in IC50 assay format free of charge.

Turnaround time: 2-3 weeks. Expedited scheduling and data delivery can be arranged prior to study commencement.

Report: A detailed report including assay conditions and comprehensive evaluation of data as well as raw data for each analysis will be provided.

Screening location: Malvern, PA, USA

Compound requirements: In brief, 50 to 100 µl of a 10 to 50 mM DMSO stock or 2-3 mg solid material is needed. Please refer to our FAQs for information regarding compound preparation and shipping.

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