Functional Assays

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.

The Chemi-Verse™ PARP7 Assay Kit is an ELISA-type chemiluminescent assay designed to measure PARP7 enzymatic activity for screening and profiling applications. PARP7 catalyzes the NAD-dependent ADP-ribosylation of histones. This 96-well or 384-well format assay kit contains sufficient amounts of purified PARP7 enzyme (amino-acids 400- 657), histone mixture, and PARP assay buffer for 100 or 400 enzyme reactions. The Chemi-Verse™ PARP7 Assay Kit takes advantage of a highly specific ADP-ribose binding reagent. First, histone proteins are coated on a 384-well plate. Next, an NAD⁺ substrate is incubated with the PARP7 enzyme in an optimized assay buffer. Finally, the plate is treated with the ADP-ribose specific binding reagent and secondary HRP-conjugated antibody, followed by addition of the ELISA ECL substrate to produce chemiluminescence that can be measured using a chemiluminescence reader. <img src="{{media url="wysiwyg/PARPs/thumbnail_HS-PARP7-diagram-large.jpg"}}" alt=""

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.