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PARP2 Homogenous Assay Kit

CAT: 0421-78572Size: 384 ReactionsDry Ice: YesHazardous: No
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CAT#:0421-78572Size:384 Reactions
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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.
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.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 activityNeed us to run inhibitor screens or profile your compounds against PARP2? Check out ourPARP/PARPTrap™ Screening ServicesorDNA Replication and Repair Screening Services.
UniProt
Q9UGN5
Applications
Screen small molecule inhibitors for drug discovery and high throughput screening (HTS) applications.
Format
Catalog #NameAmountStorage80502PARP2, GST-Tag*2 µg-80°C5X PP-01 Assay Buffer2 x 1 ml-80°CBiotinylated Histone Substrate500 reactions-80°C78311ADP-Ribose Binding Reagent 110 µl-80°C750 µM NAD+400 µl-80°C0.5 mM DTT200 µl-20°C4X Detection Buffer 1NP2 ml-20°C*The initial concentration of enzyme is lot-specific and will be indicated on the tube containing the protein.
Shipping Conditions
-80°C
Storage Conditions
This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Notes
Troubleshooting GuideVisitbpsbioscience.com/assay-kits-faqfor 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

UniProtKB · Q9UGN5

Poly [ADP-ribose] polymerase 2

PARP2_HUMAN · Homo sapiens

View on UniProt ↗
Primary accession
Q9UGN5
Review status
UniProtKB reviewed (Swiss-Prot)
Gene
PARP2
Protein existence
1: Evidence at protein level
Organism
Homo sapiens (Human)
Taxonomy ID
9606
Alternative names
PARP-2, hPARP-2
EC number
2.4.2.30
Processing
—
Secondary accessions
Q8TEU4, Q9NUV2, Q9UMR4, Q9Y6C8
Protein keywords

Technical term

3D-structureProteomics identificationReference proteome

PTM

AcetylationADP-ribosylationPhosphoprotein

Molecular function

Allosteric enzymeDNA-bindingGlycosyltransferaseNucleotidyltransferaseTransferase

Coding sequence diversity

Alternative splicing

Cellular component

ChromosomeNucleus

Biological process

DNA damageDNA repair

Ligand

NAD

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