Functional Assays

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. *NOTE: As of April 2022, this protocol has been re-optimized for performance. Previous versions of this kit are available upon request.

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. *NOTE: As of April 2022, this protocol has been re-optimized for performance. Previous versions of this kit are available upon request.

The CHK1 Kinase Assay Kit is designed to measure CHK1 kinase activity for screening and profiling applications using ADP-Glo® as a detection reagent. 

The PROTAC Optimization Kit for BRD9-Cereblon Binding is designed for the testing and profiling of PROTACs directed against BRD9 and Cereblon (CRBN). The BRD9 inhibitor BI-7273 is included as a control inhibitor of PROTAC binding to BRD9. With this kit, three simple steps are required for the measurement of PROTAC activity. First, the PROTAC of interest is incubated with CRBN and BRD9. Next, acceptor beads are added, then donor beads, followed by reading of the Alpha-counts. Illustration of the assay principle: a PROTAC of interest or positive control dBRD9 (PROTAC) interacts with both BRD9 and CRBN, bringing them in close proximity. BRD9 contains a GST tag, recognized by the GSH donor bead, while CRBN contains a FLAG tag that binds to the AlphaLISA™ acceptor bead conjugated with an anti-FLAG antibody. Upon excitation of the donor bead a singlet oxygen is generated by the donor bead, which excites the acceptor bead and emits light proportionally to the level of interaction. AlphaLISA™

The CHK2 Kinase Assay Kit is designed to measure CHK2 kinase activity for screening and profiling applications using ADP-Glo® as a detection reagent. 

The NEDD4 intrachain TR-FRET Assay Kit is a sensitive high-throughput screening (HTS) TR-FRET Assay Kit, designed to measure NEDD4 auto-ubiquitination activity in a homogeneous 384 reaction format. It utilizes a Europium-labeled ubiquitin (Ub) donor as well as Cy5-labeled Ub acceptor to complete the TR-FRET pairing. Since both the TR-FRET donor and acceptor are incorporated into poly-ubiquitin chains formed on NEDD4, this assay measures poly-ubiquitination. As a homogeneous assay, it requires no time-consuming washing steps, making it especially suitable for HTS applications as well as real-time analyses.

The ITK Kinase Assay Kit is designed to measure ITK kinase activity for screening and profiling applications using ADP-Glo^® as a detection reagent. 

The EIF2AK2 Kinase Assay Kit is designed to measure EIF2AK2 kinase activity for screening and profiling applications using ADP-Glo^® as a detection reagent. 

The PARP1 Homogeneous Assay Kit is designed to measure PARP1 activity for screening and profiling applications. PARP1 is known to catalyze the NAD-dependent addition of poly(ADP-ribose) to histones. The key to the PARP1 Homogeneous Assay Kit is a highly specific antibody that recognizes PARylated substrate. With this kit, only three simple steps are required for PARP1 reactions. First, a sample containing PARP1 enzyme is incubated with biotinylated substrate and NAD for one hour. Next, acceptor beads and primary antibody are added, then donor beads, followed by reading the Alpha-counts.