Functional Assays

The KRAS(G12V) Nucleotide Exchange Assay is a homogeneous assay designed for the screening and profiling of KRAS(G12V) antagonists/inhibitors by using BODIPY®-GDP to monitor the GDP or GTP binding status. The kit can be used with two different protocols for greater flexibility, either titrating the inhibitor at a fixed GTP concentration or titrating the GTP at a fixed inhibitor concentration. BODIPY® FL-GDP is a mixed isomer in which the fluorophore has been attached to the 2' or 3' position of the ribose ring via a linker. It is a green-fluorescent dye with similar excitation and emission to fluorescein or Alexa Fluor™ 488, characterized by a high extinction coefficient and high quantum yield and is relatively insensitive to pH changes. The dye has an excited-state lifetime of 5 nanoseconds or longer. Assay Principle Illustration: KRAS is activated upon binding GTP, when it undergoes a conformational change. KRAS then returns to a GDP-bound inactive state following the hydrolysis of G

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

The IRAK1 Kinase Assay Kit is designed to measure IRAK1 kinase activity for screening and profiling applications using ADP-Glo^® as a detection reagent.

The IRAK4 Kinase Assay Kit is designed to measure IRAK4 kinase activity for screening and profiling applications using ADP-Glo^® as a detection reagent. 

The Tropomyosin receptor kinase C (TRKC) Kinase Assay Kit is designed to measure TRKC kinase activity for screening and profiling applications using Kinase-Glo^® MAX as a detection reagent. 

The DCAF15 Intrachain TR-FRET Assay Kit is a sensitive high-throughput screening (HTS) TR-FRET Assay Kit designed to measure DCAF15 auto-ubiquitination activity in a homogeneous 384-reaction format. It utilizes Europium-labeled Ubiquitin (donor) and Cy5-labeled Ubiquitin (acceptor) to complete the TR-FRET pairing. This assay measures poly-ubiquitination since both the TR-FRET donor and acceptor are incorporated into poly-ubiquitin chains formed on DCAF15. As a homogenous assay, it requires no time-consuming washing steps, making it especially suitable for HTS applications as well as real-time kinetics analyses.

The RdRp (SARS-CoV-2) TR-FRET Assay Kit is designed to measure the activity of the SARS-CoV-2 RNA-dependent RNA Polymerase (RdRp) for screening and profiling applications. RdRp operates as a complex of NSP12, NSP7, and NSP8 proteins. As a crucial enzyme in the life cycle of coronaviruses, RdRp represents one of the most promising druggable targets for SARS-CoV-2. The assay measures the direct incorporation of biotinylated ATP into the double-stranded RNA substrate. The increase in TR-FRET signal is proportional to the amount of ATP incorporated in the RNA. With this homogeneous kit, only two steps are required for RdRp activity detection. First, a test compound is incubated with the enzyme in the reaction mixture. Next, Dye-labeled Acceptor and Eu-labeled Antibody Donor are added, followed by reading of the TR-FRET signal.

The WWP1 intrachain TR-FRET Assay Kit utilizes a Europium cryptate-labeled Ubiquitin (donor) as well as 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 WWP1, this FRET-based assay requires no time-consuming washing steps, making it especially suitable for HTS applications as well as real-time kinetic analyses of polyubiquitination.

The MURF3 intrachain TR-FRET Assay Kit utilizes a Europium cryptate-labeled 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 MURF3, this FRET-based assay requires no time-consuming washing steps, making it especially suitable for HTS applications as well as real-time kinetic analyses of polyubiquitination.