PARP2 Homogenous Assay Kit

Catalog #

78572

$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 PARP2 Homogeneous Assay Kit is designed to measure PARP2 (poly-(ADP-ribose) polymerase 2) activity for screening and profiling applications. The PARP2 Homogeneous Assay Kit comes in a convenient 384-well AlphaLISA® format, with enough purified recombinant PARP2 (amino acids 2-583), biotinylated histone substrate, ADP-Ribose Binding Reagent 1 and assay buffer for 400 enzyme reactions.

PARP2 Schematic

Figure 1: PARP2 Homogenous Assay Kit schematic.
A sample containing PARP2 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 Reagent 1, and finally donor beads. Alpha-counts are then measured. Alpha-counts are directly proportional to PARP2 activity

Need us to run inhibitor screens or profile your compounds against PARP2? Check out our PARP/PARPTrap™ Screening Services or DNA Replication and Repair Screening Services.

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Synonyms

PARP-2

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Product Data Gallery

Inhibition of PARP2 activity by Talazoparib, Olaparib 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
80502	PARP2, GST-Tag*	2 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 #

Q9UGN5

Background

PARP2 (poly(ADP-ribose polymerase 2) is part of the PARP family of proteins, involved in poly(ADP-ribosyl)ation reactions. PARP proteins, mainly PARP1 and PARP2, respond to cellular stress and are involved in DNA repair and transcriptional regulation. PARP2 is linked to oxidative stress, by altering the subcellular distribution of NRF2 (nuclear factor erythroid 2-related factor 2). Under normal conditions NRF2 forms a complex with KEAP1 (Kelch like-ECH-associated protein 1) and it is targeted for degradation by the Cul-3-Rbx1-E3 ligase complex. Under stress conditions KEAP1 releases NRF2, and NRF2 can translocate to the nucleus, a process that requires PARP2. Dysfunction in the cell redox balance and inability to repair DNA contributes to pathologies like inflammation, cancer and cardiovascular diseases. Four PARP1/2 inhibitors are currently used in the clinic for the treatment of BRCA-deficient breast, ovarian, prostate and pancreatic cancer, and the discovery of new inhibitors will lead to new avenues for the treatment of PARP linked diseases.

References

Gui B., et al., 2019 Proc Natl Acad Sci USA 116 (29): 14573-14582

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 PARP2 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