TNKS2 (PARP5B) Chemiluminescent Assay Kit

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Catalog #

78406

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Description

The TNKS2 Chemiluminescent Assay Kit is designed to measure the activity of Tankyrase 2 (TNKS2, also known as PARP5B) for screening and profiling applications. TNKS2 is known to catalyze the NAD-dependent ADP-ribosylation of histones. The key to the TNKS2 Chemiluminescent Assay Kit is the biotinylated NAD+. With this kit, only three simple steps are required for TNKS2 reactions. First, 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. Finally, the plate is treated with streptavidin-HRP followed by addition of the ELISA ECL substrate to produce chemiluminescence that can be measured using a chemiluminescence reader.

TNKS2 PARP5B Chemiluminescent

Figure 1. TNKS2 (PARP5B) Chemiluminescent Assay Kit schematic

*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 our PARP/PARPTrap™ Screening Services or DNA Replication and Repair Services.

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Synonyms

Poly [ADP-ribose] polymerase tankyrase-2, ADP-ribosyltransferase diphtheria toxin-like 6, ARTD6, Poly [ADP-ribose] polymerase 5B, Protein poly-ADP-ribosyltransferase tankyrase-2, TNKS-2, TRF1-interacting ankyrin-related ADP-ribose polymerase 2, Tankyrase,

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Product Data Gallery

Product Info

Storage and Usage

Citations

Assay Kit Format

Chemiluminescent

Supplied As

The TNKS2 assay kit comes in a convenient 96-well or 384-well format, with purified TNKS2 enzyme, histone mixture, and PARP assay buffer for 100 or 400 enzyme reactions.

Materials Required But Not Supplied

DTT (10 mM in water, prepared fresh)
1x PBS (phosphate buffer saline) buffer
PBST buffer (1x PBS, containing 0.05% Tween-20)
Luminometer or microplate reader capable of reading chemiluminescence
Adjustable micropipettor and sterile tips
Rotating or rocker platform

Format

96 Reactions

Catalog #	Name	Amount	Storage
80515	TNKS2* (Tankyrase 2 (PARP5B) [849-1166], GST-tag)	1 µg	-80°C	Avoid freeze/ thaw cycles
52029	5x histone mixture	1 ml	-80°C
78371	PARP Substrate Mixture 2	250 µl	-80°C
80602	10x PARP assay buffer	1 ml	-20°C
79743	Blocking buffer 3	25 ml	+4°C
80611	Streptavidin-HRP	100 µl	+4°C
79670	ELISA ECL Substrate A (translucent bottle)	6 ml	Room Temp
79670	ELISA ECL Substrate B (brown bottle)	6 ml	Room Temp
79837	96-well module plate		Room 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 Reactions

Catalog #	Name	Amount	Storage
80515	TNKS2*	2 µg	-80°C	Avoid freeze/ thaw cycles
52029	5x histone mixture	2 x 1 ml	-80°C
78371	PARP Substrate Mixture 2	2 x 250 µl	-80°C
80602	10x PARP assay buffer	2 x 1 ml	-20°C
79743	Blocking buffer 3	2 x 25 ml	+4°C
80611	Streptavidin-HRP	2 x 100 µl	+4°C
79670	ELISA ECL Substrate A (translucent bottle)	2 x 6 ml	Room Temp
79670	ELISA ECL Substrate B (brown bottle)	2 x 6 ml	Room Temp
78188	384-well plate		Room Temp

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

UniProt #

Q9H2K2

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

Study enzyme kinetics and screen small molecular inhibitors for drug discovery and high throughput (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 TNKS2 Chemiluminescent Assay Kit is compatible with up to 1% final DMSO concentration. We recommend preparing the inhibitor in no higher than 10% DMSO solution in buffer and using 5 µl per well.

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