Assays & Kits

The USP20 Inhibitor Screening Assay Kit is a 96-well format fluorogenic assay designed to measure the activity of the deubiquitinating (DUB) enzyme USP20 for screening and profiling applications. To determine the effect of an inhibitor on USP20 activity, the enzyme should be preincubated with or without the test inhibitor prior to adding the Ub-AMC substrate to the reaction. The assay was functionally validated using Ub-Aldehyde, a potent inhibitor of DUB subfamilies Ubiquitin C-terminal Hydrolases (UCHs), Ubiquitin-Specific Proteases (USPs), Ovarian Tumor Proteases (OTU), and Machado-Josephin Domain (MJD) proteases. <img src="{{media url="wysiwyg/ubiquitin/78831.png"}}" alt="" width="500" height="226" /> Figure 1: Illustration of the assay principle. <br />Ubiquitin-AMC is a fluorogenic substrate for ubiquitin hydrolases based on the C-terminus derivatization of ubiquitin with 7-amido-4-methylcoumarin (AMC). In the conjugated form, the energy emitted from fluorochrome AMC is quenche

The MDM2-Driven p53 Ubiquitination Assay kit is a sensitive AlphaLisa® high-throughput screening (HTS) assay designed to measure MDM2 (mouse double minute 2 homolog) E3 ligase activity in a homogeneous 384 reaction format. The assay kit comes with enough biotinylated ubiquitin, ATP, FLAG-tagged p53, assay buffer, detection buffer, purified UBE1 (E1), UbcH5b (E2), and MDM2 (E3) for 384 reactions. The assay can detect mono-ubiquitination and poly-ubiquitination of p53. E1 and E2 enzymes are incubated with the E3 complex and FLAG-tagged p53, in the presence of biotin-conjugated ubiquitin and ATP. Ubiquitination of p53 occurs in a multistep ubiquitin transfer from E1 to E2 to E3, and E3-mediated conjugation of ubiquitin to p53. Next, acceptor beads are added, followed by streptavidin-conjugated donor beads. Alpha-counts are then measured. The increase in Alpha-counts is proportional to the mono- or poly-ubiquitination of the FLAG-tagged p53.

The Chemi-Verse™ PAK4 Kinase Assay Kit is designed to measure PAK4 (p21-activated kinase 4) serine/threonine kinase activity for screening and profiling applications using ADP-Glo™ as a detection reagent. The assay kit comes in a convenient 96-well format, with enough purified recombinant PAK4 kinase, kinase substrate, ATP, and kinase assay buffer for 100 enzyme reactions.

The Chemi-Verse™ PAK6 Kinase Assay Kit is designed to measure PAK6 (p21-activated kinase 6) serine/threonine kinase activity for screening and profiling applications using ADP-Glo™ as a detection reagent. The assay kit comes in a convenient 96-well format, with enough purified recombinant PAK6 kinase, kinase substrate, ATP, and kinase assay buffer for 100 enzyme reactions.

The Chemi-Verse™ PAK7 Kinase Assay Kit is designed to measure PAK7 (p21-activated kinase 7) serine/threonine kinase activity for screening and profiling applications using ADP-Glo™ as a detection reagent. The assay kit comes in a convenient 96-well format, with enough purified recombinant PAK7 kinase, kinase substrate, ATP, and kinase assay buffer for 100 enzyme reactions.

Measure 3CL protease (3CLpro) inhibition or screen for inhibitors with the live cell, fluorescent 3CLglow assay. The 3CLglow sensor expresses a green fluorescent protein in the presences of the 3CLpro enzyme as well as a constitutively expressed red fluorescent protein to monitor cell health and compound toxicity. The assay is packaged in BacMam for expression in mammalian cells. BacMam provides even, near 100% transduction efficiency in HEK293 cells as well as expression in many other cell lines, iPSCs, and primary cultures including A549s and HUVECs. Expression is easily titrateable to your experimental needs or to adjust assay sensitivity. This kit includes the 3CLglow sensor BacMam, 3CLpro enyme BacMam, and 500mM sodium butyrate in H2O. 5mL and 30mL sizes are also available.

Measure real-time cAMP signaling kinetics in the primary cilia with this live cell fluorescent assay. Add 5HT6-tagged cADDis to your cells and measure robust signals the next day on a fluorescence imaging system. Decreasing green fluorescence indicates ciliary cAMP increase. This assay is genetically encoded and packaged in BacMam for efficient transduction in most cell types - including uniform and near 100% efficiency in HEK293 cells as well as expression in many primary or iPSC-derived cells. The kit includes the cilia-targeted cAMP sensor BacMam and 500mM sodium butyrate in H2O. 10mL size is available as well.

Measure real-time cAMP signaling kinetics in the primary cilia with this live cell fluorescent assay. Add 5HT6-tagged cADDis to your cells and measure robust signals the next day on a fluorescence imaging system. Decreasing green fluorescence indicates ciliary cAMP increase. Constitutively expressed cytosolic red fluorescence can be used for ratiometric measurements. This assay is genetically encoded and packaged in BacMam for efficient transduction in most cell types - including uniform and near 100% efficiency in HEK293 cells as well as expression in many primary or iPSC-derived cells. The kit includes the cilia-targeted cAMP sensor BacMam and 500mM sodium butyrate in H2O. 30mL size is available as well.

Measure real-time cAMP signaling kinetics in the primary cilia with this live cell fluorescent assay. Add 5HT6-tagged cADDis to your cells and measure robust signals the next day on a fluorescence imaging system. Decreasing green fluorescence indicates ciliary cAMP increase. Constitutively expressed cytosolic red fluorescence can be used for ratiometric measurements. This assay is genetically encoded and packaged in BacMam for efficient transduction in most cell types - including uniform and near 100% efficiency in HEK293 cells as well as expression in many primary or iPSC-derived cells. The kit includes the cilia-targeted cAMP sensor BacMam and 500mM sodium butyrate in H2O. 10mL size is available as well.