Functional Assays

The Tie2 Kinase Assay Kit is designed to measure Tie2 kinase activity for screening and profiling applications using ADP-Glo® as a detection reagent. 

The TNKS1 Colorimetric Assay Kit is designed to measure the activity of Tankyrase 1 (TNKS1, also known as PARP5A) for screening and profiling applications. TNKS1 is known to catalyze the NAD-dependent ADP-ribosylation of histones. The key to the TNKS1 (PARP5A) Colorimetric Assay Kit is the biotinylated NAD+. With this kit, only three simple steps are required for TNKS1 reactions. First, histone proteins are coated on a 96-well plate. Next, a biotinylated NAD+ mix (termed PARP Substrate Mixture) is incubated with the TNKS1 enzyme in an optimized assay buffer. Finally, the plate is treated with streptavidin-HRP followed by addition of the colorimetric HRP substrate to produce color that can be measured using a UV/Vis spectrophotometer microplate reader.

The TNKS2 Colorimetric 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 Colorimetric 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 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 Colorimetric HRP substrate to produce color that can be measured using a UV/Vis spectrophotometer microplate reader.

The IL-2: IL-2 Receptor Alpha (IL-2RA) Inhibitor Screening Colorimetric Assay Kit is designed for the screening and profiling of inhibitors or neutralizing antibodies blocking the interaction between human IL-2 and its receptor IL-2RA.  The assay requires only a few steps. First, IL-2RA is coated on a 96-well plate overnight. After blocking, the receptor is pre-incubated with the test inhibitor or neutralizing antibody. Upon subsequent incubation with Biotin-IL-2, the plate is treated with Streptavidin-HRP followed by addition of a colorimetric HRP substrate to produce color, which can be quenched and measured using a UV/Vis microplate reader.

The ChoosE3-Freedom Intrachain TR-FRET Assay Kit contains E1 and E2 enzymes, ATP, an optimized TRF Ubiquitin Mix, and a universal buffer. Purified E3 ligase MDM2 is also provided as an internal quality control. The assay was designed with a Europium-labeled Ubiquitin donor and a Cy5-labeled Ubiquitin acceptor to complete the TR-FRET pairing. Since both the TR-FRET donor and acceptor are incorporated into poly-ubiquitin chains formed on the E3 ligase, the assay measures only poly-ubiquitination and not mono-ubiquitination. This FRET-based assay requires no time-consuming washing steps, making it especially suitable for HTS applications as well as real-time kinetics.

ChoosE2-Opti™ Intrachain TR-FRET Assay Kit is a sensitive high-throughput screening (HTS) TR-FRET Assay Kit, designed to identify E2 enzyme(s) that are functional partners for a purified E3 ubiquitin ligase of interest and lead to its self-polyubiquitination. The assay kit contains five E2 enzymes to be tested with the E3 ligase of interest, and two E3 ligases (SMURF1 and MDM2) to use as internal controls.  It also contains an E1 enzyme, ATP, an optimized TRF Ubiquitin Mix, and a universal buffer.  It utilizes a Europium-labeled Ubiquitin Donor as well as Cy5-labeled Ubiquitin Acceptor to complete the TR-FRET pairing. Since both the TR-FRET donor and acceptor are incorporated into polyubiquitin chains, this FRET-based assay requires no time-consuming washing steps, making it especially suitable for HTS applications as well as real-time kinetics analyses.

The KRAS(G12C) Coupled Nucleotide Exchange Assay Kit is designed for screening and profiling of KRAS(G12C) antagonists/inhibitors by monitoring the binding of an effector protein such as the Ras binding domain of Raf1, (RBD-cRaf) to KRAS(G12C). With this kit, a few simple steps on a microtiter plate are required for nucleotide exchange detection. First, a sample containing GDP-loaded KRAS(G12C) is incubated with SOS1 and GTP for the nucleotide exchange. Next, RBD-cRAF is added and incubated for the effector-RAS binding. Then, acceptor and donor beads are added and incubated for detection followed by reading the Alpha-counts. SOS1 (son of sevenless) is a guanine nucleotide exchange factor that facilitates the exchange of GDP for GTP. GDP-loaded KRAS(G12C) is in an inactive state, and does not interact with the Ras-binding domain (RBD) of cRAF.  SOS1 assists in the release of GDP from KRAS(G12C) so that GTP can occupy the nucleotide binding pocket. This results in a conformational change

The KRAS(G12D) Coupled Nucleotide Exchange Assay Kit is designed for screening and profiling of KRAS(G12D) antagonists/inhibitors by monitoring the binding of an effector protein such as the Ras binding domain of Raf1, (RBD-cRaf) to KRAS(G12D). With this kit, a few simple steps on a microtiter plate are required for nucleotide exchange detection. First, a sample containing GDP-loaded KRAS(G12D) is incubated with SOS1 and GTP for the nucleotide exchange. Next, RBD-cRAF is added and incubated for the effector-RAS binding. Then, acceptor and donor beads are added and incubated for detection followed by reading the Alpha-counts. SOS1 (son of sevenless) is a guanine nucleotide exchange factor that facilitates the exchange of GDP for GTP. GDP-loaded KRAS(G12D) is in an inactive state and does not interact with the Ras-binding domain (RBD) of cRAF.  SOS1 assists in the release of GDP from KRAS(G12D) so that GTP can occupy the nucleotide binding pocket. This results in a conformational change

Please note this product may be subject to fees, we invite you to contact your local office. The KRAS IsoformB Coupled Nucleotide Exchange Assay Kit is designed for screening and profiling of KRAS IsoformB antagonists/inhibitors by monitoring the binding of an effector protein such as the Ras binding domain of Raf1, (RBD-cRaf) to KRAS. With this kit, a few simple steps on a microtiter plate are required for nucleotide exchange detection. First, a sample containing GDP-loaded KRAS Isoform B is incubated with SOS1 and GTP for the nucleotide exchange. Next, RBD-cRAF is added and incubated for the <br />effector-RAS binding. Then, acceptor and donor beads are added and incubated for detection followed by reading the Alpha-counts. SOS1(son of sevenless)is a guanine nucleotide exchange factor that facilitates the exchange of GDP for GTP. GDP-loaded KRAS Isoform B is in an inactive state and does not interact with the Ras-binding domain (RBD) of cRAF. SOS1 assists in the release of GDP from K