TNKS2 (PARP5B) Chemiluminescent Assay Kit
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.
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.
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- 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
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 | |
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 | |
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.