Functional Assays

The Fibroblast growth factor receptor 4 (FGFR4) Kinase Assay Kit is designed to measure FGFR4 kinase activity for screening and profiling applications using ADP-Glo® as a detection reagent. 

The C-terminal Src kinase (CSK) Kinase Assay Kit is designed to measure CSK kinase activity for screening and profiling applications using ADP-Glo® as a detection reagent. 

The transforming growth factor beta receptor (TGFβR1, also known as ALK5) Kinase Assay Kit is designed to measure the serine/threonine activity of TGFβR1 (ALK5) for screening and profiling applications using ADP-Glo^® as a detection reagent. 

The CBL-B-driven SRC ubiquitination intrachain TR-FRET Assay Kit utilizes a Europium cryptate-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, this assay does not detect mono-ubiquitination. The FRET-based format requires no time-consuming washing steps, making it especially suitable for HTS applications as well as real-time analyses of polyubiquitination. <img style="display: block; margin-left: auto; margin-right: auto;" src="{{media url="wysiwyg/ubiquitin/78820.png"}}" alt="" width="900" height="220" /> Figure 1: CBL-B-driven SRC ubiquitination intrachain TR-FRET Assay Kit schematic.

The CBL-B-driven AXL ubiquitination intrachain TR-FRET Assay Kit utilizes a Europium cryptate-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, this assay does not detect mono-ubiquitination. The FRET-based format requires no time-consuming washing steps, making it especially suitable for HTS applications as well as real-time analyses of polyubiquitination. <img src="{{media url="wysiwyg/ubiquitin/78821.png"}}" alt="" width="850" height="207" /> Figure 1: CBL-B-driven AXL ubiquitination intrachain TR-FRET Assay Kit schematic.

The C-CBL-driven SRC Ubiquitination Intrachain TR-FRET Assay Kit utilizes a Europium cryptate-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, this assay does not detect mono-ubiquitination. The FRET-based format requires no time-consuming washing steps, making it especially suitable for HTS applications as well as real-time analyses of polyubiquitination. <img src="{{media url="wysiwyg/ubiquitin/78822.png"}}" alt="" width="850" height="207" /> Figure 1: C-CBL-driven SRC ubiquitination intrachain TR-FRET Assay Kit schematic.

The C-CBL-driven AXL Ubiquitination Intrachain TR-FRET Assay Kit utilizes a Europium cryptate-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, this assay does not detect mono-ubiquitination. The FRET-based assay requires no time-consuming washing steps, making it especially suitable for HTS applications as well as real-time analyses of polyubiquitination. <img src="{{media url="wysiwyg/ubiquitin/78823.png"}}" alt="" width="850" height="207" /> Figure 1: C-CBL-driven AXL ubiquitination intrachain TR-FRET Assay Kit schematic.

The FGL1:LAG3 Inhibitor Screening Assay is an AlphaLISA®-based assay designed to measure the inhibition of LAG3 binding to FGL1 in a homogeneous 384-reaction format. The assay is straightforward: first, FGL1 and LAG3 are incubated with or without a compound of interest. Then, acceptor and donor beads are added to the reaction, followed by reading the Alpha-counts. <img src="{{media url="wysiwyg/Imtx/thumbnail_78824_LAG3-LGL1_illustration.png"}}" alt="" width="900" height="253" /> Illustration of the assay principle.<br />FGL1 contains a His tag recognized by the Nickel chelate acceptor bead, whereas LAG3 is biotinylated, allowing its binding to the streptavidin-coated acceptor bead. Interaction between FGL1 and its receptor LAG3 brings the donor and acceptor beads in proximity. A singlet oxygen generated upon excitation of the donor bead leads to the excitation of the acceptor bead, which emits light. Light emission in the assay is proportional to the level of interaction. AlphaLISA™

The KRAS(G12V) Isoform B Coupled Nucleotide Exchange Assay Kit is designed for screening and profiling of antagonists/inhibitors of KRAS(G12V), Isoform B, by monitoring the binding of an effector protein such as the Ras binding domain of Raf1 (RBD-cRaf) to KRAS. First, a sample containing GDP-loaded KRAS(G12V) Isoform B 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(G12V) 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 KRAS(G12V) Isoform B so that GTP can occupy the nucleotide binding pocket. This results in a conformational change in KRAS that permits its binding to RBD-cRAF. The KRAS