PARP1 Homogenous Assay Kit

Catalog #

78438

$1,470 *

Size: 384 reactions

Qty

Bulk/Custom

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*US Pricing only. For international pricing, please contact your local distributor.

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Description

The PARP1 Homogeneous Assay Kit is designed to measure PARP1 (poly-(ADP-ribose) polymerase 1) activity for screening and profiling applications. The PARP1 Homogeneous Assay Kit comes in a convenient 384-well AlphaLISA® format, with enough purified recombinant PARP1, biotinylated histone substrate, ADP-Ribose Binding Reagent 1 and assay buffer for 400 enzyme reactions.

Figure 1: PARP1 Homogenous Assay Kit schematic.
A sample containing PARP1 is incubated with a biotinylated histone substrate and NAD⁺ for one hour. This is followed by the addition of acceptor beads and ADP-ribose binding agent, and finally donor beads. Alpha-counts are then measured. Alpha-counts are directly proportional to PARP1 activity.

Need us to run inhibitor screens or profile your compounds against PARP1? Check out our PARP/PARPTrap™ Screening Services or DNA Replication and Repair Screening Services.

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Synonyms

Poly [ADP-ribose] polymerase 1, ADP-ribosyltransferase diphtheria toxin-like 1, ARTD1, DNA ADP-ribosyltransferase PARP1, NAD(+) ADP-ribosyltransferase 1, ADPRT 1, Poly[ADP-ribose] synthase 1, Protein poly-ADP-ribosyltransferase PARP1, PARP-1

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Product Data Gallery

Inhibition of PARP1 activity by Olaparib, Talazoparib and AZD-5305

Product Info

Storage and Usage

Citations

Assay Kit Format

AlphaLISA®

Materials Required But Not Supplied

AlphaLISA® Anti-Rabbit IgG Acceptor Beads (Perkin Elmer #AL104C)
AlphaScreen® Streptavidin-Conjugated Donor Beads (Perkin Elmer #6760002S)
Optiplate - 384 (Perkin Elmer #6007290)
AlphaScreen microplate reader

Format

Catalog #	Name	Amount	Storage
80501	PARP1, GST-Tag*	0.4 µg	-80°C
	5X PP-01 Assay Buffer	2 x 1 ml	-80°C
	Biotinylated Histone Substrate	500 reactions	-80°C
78311	ADP-Ribose Binding Reagent 1	10 µl	-80°C
	750 µM NAD⁺	400 µl	-80°C
	0.5 mM DTT	200 µl	-20°C
	4X Detection Buffer 1NP	2 ml	-20°C

*The initial concentration of enzyme is lot-specific and will be indicated on the tube containing the protein.

UniProt #

P09874

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.

References

Marques M., et al., 2019 Oncogene 38 (12): 2177-2191.

Storage/Stability

This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.

Applications

Screen small molecule inhibitors for drug discovery and high throughput screening (HTS) applications.

Shipping Temperature

-80°C

Notes

Troubleshooting Guide
Visit bpsbioscience.com/assay-kits-faq for detailed troubleshooting instructions. For all further questions, please email [email protected]

Contraindications

The PARP1 Homogenous Assay Kit is compatible with up to 1% final DMSO concentration.
Avoid green and blue dyes that absorb light in the AlphaScreen signal emission range (λ=520-620 nm), such as Trypan Blue.
Avoid the use of potent singlet oxygen quenchers such as sodium azide (NaN₃) or metal ions (Fe²⁺, Fe³⁺, Cu²⁺, Zn²⁺ and Ni²⁺).
The presence of >1% RPMI 1640 culture medium leads to a signal reduction due to the presence of excess biotin and iron in this medium. Media like MEM, which lacks these components, does not affect AlphaScreen assays.

Warnings

Avoid freeze/thaw cycles

Disclaimers and Limitations

BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.

End-users should immediately report any issues to [email protected] upon receipt of the kit, such as missing components, damaged vials, no dry ice, etc.

Scientific Category

PARP

Regulatory Status

Research Use Only