Products from BPS Bioscience require a minimum order value above 400€Applications: Screen molecules that bind to PARP1 in the same binding site as Olaparib for drug discovery high-throughput screening (HTS) applications.Determine IC
50 values of PARP1 inhibitors with the same binding site as Olaparib.
Background: PARP1, also known as poly-(ADP-ribose) polymerase 1 or NAD
+ ADP-ribosyltransferase 1, is part of the PARP family, and it is the most abundant member. ADP ribosylation, which is the addition of an ADP-ribose to a protein, is a reversible post-translational modification of proteins mostly involved in the DNA Damage Response (DDR) pathway. Poly-ADP-ribosylation (termed PARylation) is the addition of linear or branched chains of ADP-ribose. PARP1 participates in DNA repair by non-homologous end joining (NHEJ), homologous recombination (HR), microhomology-mediated end-joining (MMEJ) and nucleotide excision repair. Dysfunction of DDR pathways can lead to oncogenesis. Overexpression of PARP1 has been found in breast and colon cancer, neuroblastoma, and others. This overexpression can lead to increasing MMEJ, an error-prone DNA repair mechanism, and genome instability leading to cancer. In addition to being involved in DDR, PARP1 is also linked to inflammation and type I diabetes. PARP1 inhibitors have been used in cancer treatment with success. In addition to reducing MMEJ, the use of PARP1 inhibitors can lead to synthetic lethality when homologous recombination repair (HRR) mechanisms are already defective, as in the case of BRCA1 (breast cancer susceptibility protein type 1) and BRCA2 deficient cells. Further understanding of the molecular pathways involving PARP1, and this contribution to disease, will continue to pave the way for new therapies for PARP1-linked diseases.
Contraindications: This assay kit is compatible with up to 1% final DMSO concentration.Fluorescent compounds that have λex=485/20 nm and detection at λem=528/20 nm can interfere with the readouts.
Description: The PARP1 Olaparib Competitive Inhibitor Assay Kit is a competitive FP (fluorescent polarization) assay designed to measure the formation of a complex between PARP1 (poly(ADP-ribose) polymerase 1) and a fluorescent probe that contains the PARP1 inhibitor Olaparib. When the probe is bound to PARP1, FP is high. In the presence of a test compound able to bind to the same site in PARP1 as Olaparib, the Olaparib-containing fluorescent probe is displaced from PARP1 and remains in solution, resulting in low FP. The PARP1 Olaparib Competitive Inhibitor Assay Kit comes in a convenient 96-well format, with purified PARP1 enzyme, Olaparib-containing fluorescent-labeled probe, and assay buffer for 100 enzyme reactions.Note: This kit is not appropriate for inhibitors expected to bind to PARP1 at different sites from Olaparib.PARP1 binds to the Olaparib-containing fluorescent probe, forming a complex. This complex, when subjected to polarized excitation light, emits highly polarized light due to its restricted movement in solution. In the presence of a test compound (A), PARP1 may form a complex with either the test compound, if the compound has the same binding site of PARP1 as Olaparib, or with the Olaparib-containing fluorescent probe. If the test compound binds to PARP1 at the same site, the Olaparib-containing fluorescent is freed and a decrease in FP is observed. The decrease in FP value is proportional to the competitive binding of the test compound to PARP1.This assay requires a fluorescent microplate reader capable of measuring fluorescence polarization (FP) to read the FP signal. For more information FP technology, visit our Tech Note:
FP, assay principles and applications.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Warnings: This assay requires a fluorescent microplate reader capable of measuring fluorescence polarization (FP) and equipped with the required parts to read the FP signal. For more information FP technology, visit our Tech Note: "FP, assay principles and applications", https://bpsbioscience.com/fluorescence-polarization-assays-principles-applications Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
References: Marques M., et al., 2019 Oncogene 38 (12): 2177-2191.
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