TNKS2 (PARP5B) Chemiluminescent Assay Kit

The TNKS2 (PARP5B) Chemiluminescent Assay Kit is designed to measure the activity of TNKS2 (Tankyrase 2) for screening and profiling applications. The TNKS2 assay kit comes in a convenient 96-well or 384-well format, with enough recombinant purified TNKS2 enzyme (amino acids 849-1166), reaction substrates, and PARP assay buffer for 100 or 400 enzyme reactions.Figure 1. TNKS2 (PARP5B) Chemiluminescent Assay Kit schematic.Histone proteins are coated on a 96-well or 384-well plate. Next, a biotinylated NAD+mix (termed PARP Substrate Mixture) is incubated with the TNKS2 enzyme in an optimized assay buffer. The plate is then treated with streptavidin-HRP followed by addition of the ELISA ECL substrate to produce chemiluminescence that can be measured using a chemiluminescence reader. The chemiluminescence signal is proportional to the TNKS2 activity.*NOTE: As of April 2022, this protocol has been re-optimized for performance. Previous versions of this kit are available upon request.Need us to run inhibitor screens or profile your compounds against TNKS2 (PARP5B) ? Check out ourPARP/PARPTrap™ Screening ServicesorDNA Replication and Repair Services.

Product Specifications

Background

TNKS2 (Tankyrase 2), also known as poly- (ADP-ribose) polymerase 5b (PARP5B), is part of the PARP family. 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 units of ADP-ribose. TNKS2 is involved in Wnt/β-catenin and Hippo signaling, telomere maintenance, DNA damage repair and glucose balance. TNKS2 acts on several substrates, including PTEN (phosphatidylinositol (3,4,5) -triphosphase phosphatase), Axin1/2 and RNF46 (RING finger protein 146) . Dysfunction of its activity can result in obesity, diabetes, fibrosis, cherubism and cancer. TNKS2 is found at high levels in colon, lung, brain, breast cancer, amongst others. The development and use of TNKS2 inhibitors has been an active and exciting research area. In colorectal cancer, one of the most common cancers worldwide, the Wnt/β-catenin pathway is found to be hyperactivated. The use of TNKS2 inhibitors resulted in decreased cell proliferation by stabilizing the levels of Axin. XAV-939, a TNKS inhibitor, has been tested in ovarian and breast cancer, neuroblastoma, and hepatocellular carcinoma, resulting in decreased cell proliferation and metastasis and increased apoptosis. The use of TNKS2 inhibitors can also sensitize cells to treatment with immune checkpoint inhibitors, as in the case of combination with anti-PD-1 (programmed cell death protein 1) in melanoma. A deeper knowledge of the functions of TNKS2 and its partners will broaden our knowledge of this protein, and how best to take advantage of its properties in the clinic.

UniProt

Q9H2K2

Applications

Study enzyme kinetics and screen small molecule inhibitors for drug discovery and high throughput screening (HTS) applications.

Format

96 ReactionsCatalog #NameAmountStorage80515Tankyrase 2 (PARP5B) [849-1166], GST-Tag*1 µg-80°C520295x Histone Mixture1 ml-80°C78371PARP Substrate Mixture 2250 µl-80°C8060210x PARP Assay Buffer1 ml-20°C79743Blocking Buffer 325 ml+4°C0.5 M DTT200 µl-20°C80611Streptavidin-HRP100 µl+4°C79670ELISA ECL Substrate A (translucent bottle) 6 mlRoom TempELISA ECL Substrate B (brown bottle) 6 mlRoom Temp7983796-well module plate1Room Temp*The concentration of the protein is lot-specific and will be indicated on the tube. Excess material has been given for ease of retrieval.384 ReactionsCatalog #NameAmountStorage80515Tankyrase 2 (PARP5B) [849-1166], GST-Tag*2 µg-80°C520295x Histone Mixture2 x 1 ml-80°C78371PARP Substrate Mixture 22 x 250 µl-80°C8060210x PARP Assay Buffer2 x 1 ml-20°C79743Blocking Buffer 32 x 25 ml+4°C0.5 M DTT2 x 200 µl-20°C80611Streptavidin-HRP2 x 100 µl+4°C79670ELISA ECL Substrate A (translucent bottle) 2 x 6 mlRoom TempELISA ECL Substrate B (brown bottle) 2 x 6 mlRoom Temp78188384-well plate1Room Temp*The concentration of the protein is lot-specific and will be indicated on the tube.

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]