Functional Assays

The PDE3A TR-FRET Assay Kit is designed to provide fast and easy identification of inhibitors of PDE3A (phosphodiesterase 3A) using Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET). The PDE3A TR-FRET Assay Kit comes in a convenient 96-well or 384-well format, with enough recombinant purified PDE3A (amino acids 669-end) enzyme, fluorescently labeled PDE substrate (cAMP), binding agent, and PDE assay buffer for 100 or 384 enzyme reactions. The reaction uses a fluorescein-conjugated (FAM) cyclic monophosphate nucleotide.  Phosphodiesterase PDE3A catalyzes the hydrolysis of the phosphodiester bond in the cyclic monophosphate nucleotide, releasing the phosphate group for binding. The phosphate group binds to a "Binding Agent" (BA) that is recognized by terbium-labeled donor beads. This results in energy transfer from the terbium to FAM, which emits a fluorescent signal at 520 nm. If unbound to the phosphate group, the terbium-labeled beads emit at λ=490 nm. The fluorescent int

The SARM1 Fluorogenic Assay Kit (Hydrolase Activity) is designed to measure NAD⁺ cleavage activity in screening and profiling applications. The SARM1 assay kit comes in a convenient 96-well or 384-well format, with enough recombinant human SARM1 enzyme, its substrate N6-etheno-NAD (e-NAD), and SARM1 assay buffer for 100 or 400 enzyme reactions. In addition, the kit includes a SARM1 inhibitor (DSRM-3716) for use as a control inhibitor. Hydrolase activity of SARM1 is measured by following the hydrolysis of Etheno-NAD to form Etheno-ADPR + nicotinamide. Etheno-NAD is not fluorescent due to internal quenching. Upon hydrolysis by SARM1, nicotinamide is released, leading to an increase in fluorescence signal directly proportional to the enzymatic activity.

The DCAF11 Intrachain TR-FRET Assay Kit is a sensitive high-throughput screening (HTS) TR-FRET Assay Kit designed to measure DCAF11 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 DCAF11. As a homogenous assay, it requires no time-consuming washing steps, making it especially suitable for HTS applications as well as real-time analyses. <img style="display: block; margin-left: auto; margin-right: auto;" src="{{media url="wysiwyg/ubiquitin/78542.png"}}" alt="" width="1059" height="225" /> Figure 1. DCAF11 Intrachain TR-FRET Assay Kit Schematic.

The EPHA4 Kinase Assay Kit is designed to measure EPHA4 kinase activity for screening and profiling applications using ADP-Glo® as a detection reagent. 

The Von Hippel-Lindau tumor suppressor (VHL) Binding Assay Kit is a homogeneous assay which measures the binding of VHL to the high-affinity VHL fluorescent probe (BDY FL VH032) using Fluorescence Polarization (FP). In addition, the kit includes the VHL inhibitor VH298 for use as an inhibitor control. This assay takes advantage of the fluorescent substrate BDY FL VH032. Using this kit, only one step on a 96-well plate is required. The BDY FL VH032 is incubated with the VHL complex in the presence of a compound of interest to produce a change in fluorescent polarization. The FP signal is measured using a fluorescent microplate reader capable of measuring fluorescence polarization.

The P70S6K (S6K1) Kinase Assay Kit is designed to measure P70S6K (S6K1) kinase activity for screening and profiling applications using ADP-Glo® as a detection reagent. 

The PCSK9-LDLR Homogeneous Assay Kit is designed for screening and profiling purposes. Two pre-formulated assay buffers are supplied with this kit to validate PCSK9-LDLR binding affinity in either neutral or acidic binding conditions. With this kit, two simple steps are required for binding measurement. First, PCSK9 enzyme is incubated with LDLR in the preferred buffer for one hour. Next, donor and acceptor beads are added, followed by reading the Alpha-counts. <img src="{{media url="wysiwyg/Cholesterol/78812.png"}}" alt="" width="796" height="246" /> Figure 1: Illustration of the assay principle. The FLAG-tag of LDLR binds to the anti-FLAG acceptor beads, while the His-tag on PCSK9 binds to Nickel Chelate donor beads, bringing the acceptor and donor beads in close proximity. 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™ immunoassays are a no-w

The PCSK9(His)-LDLR Binding Assay Kit is an ELISA-type 96-well format assay designed for screening and profiling purposes. Moreover, two pre-formulated assay buffers are supplied to validate PCSK9-LDLR binding affinity in either neutral or acidic binding conditions. The assay takes advantage of the high sensitivity of detection of His-tag PCSK9 by Anti-His HRP-antibody. First, the LDLR ectodomain is coated on a 96-well plate. Next, PCSK9 is incubated with LDLR on the plate. Finally, the plate is treated with Anti-His HRP-antibody followed by addition of an HRP substrate to produce chemiluminescence, measured using a chemiluminescence reader.   Figure 1: Illustration of the ELISA-based PCSK9(His)-LDLR Binding Assay Kit.

The PCSK9(His)-LDLR Binding Assay Kit is an ELISA-type 96-well format assay designed for screening and profiling purposes. Moreover, two pre-formulated assay buffers are supplied to validate PCSK9-LDLR binding affinity in either neutral or acidic binding conditions. The assay takes advantage of the high sensitivity of detection of His-tag PCSK9 by Anti-His HRP-antibody. First, the LDLR ectodomain is coated on a 96-well plate. Next, PCSK9 is incubated with LDLR on the plate. Finally, the plate is treated with Anti-His HRP-antibody followed by addition of an HRP substrate to produce chemiluminescence, measured using a chemiluminescence reader.   Figure 1: Illustration of the ELISA-based PCSK9(His)-LDLR Binding Assay Kit.