NEDD4 intrachain TR-FRET Assay Kit | 78428

NEDD4 intrachain TR-FRET Assay Kit | 78428 | Gentaur US, UK & Europe Disrtribition

Category: Ubiquitination/Assay Kit

Application: Screen molecules that inhibit NEDD4 Ub ligase activity in drug discovery HTS applications Determine compound IC₅₀ Perform NEDD4 real-time kinetics

Background: Covalent conjugation to ubiquitin (Ub) is one of the major post-translational modifications that regulates protein stability, function, and localization. Ubiquitination is the concerted action of three enzymes: a Ub-activating enzyme (E1),  a Ub-conjugating enzyme (E2), and a Ub ligase (E3). The specificity and efficiency of ubiquitination are largely determined by the E3 enzyme, which directs the last step of the Ub-conjugating cascade by binding to both an E2~Ub conjugate and a substrate protein. This step ensures the transfer of Ub from E2~Ub to the substrate, leading to its mono- or poly-ubiquitination. NEDD4 (neural precursor cell expressed developmentally down-regulated protein 4) is an E3 ligase, member of the HECT (Homologous to the E6-AP carboxyl terminus) ubiquitin ligase family which target client proteins to the proteasome system for degradation. The protein contains an N-terminal calcium and phospholipid-binding C2 domain, three tryptophan-rich WW domains (that bind to proline-rich peptide motifs), and a C-terminal HECT catalytic domain. NEDD4 regulates the expression levels of various receptor tyrosine kinases, notably growth factor receptors IGF1R (Insulin-like growth factor 1 receptor), FGFR1 (fibroblast growth factor receptor 1), EGFR (Epidermal growth factor receptor), and VEGFR2 (Vascular endothelial growth factor receptor 2), thereby playing a role in growth factor signaling and regulating cell proliferation. The E3 ligase also controls the expression levels of ion channels and is part of a signaling complex involved in dendrite extension and neuron architecture. It is an essential protein during development, including neural development. NEDD4 is a potential therapeutic target for the treatment of various types of cancer, cardiovascular disease, and neuro-degenerative diseases such as Parkinson's disease, Alzheimer's disease or Amyotrophic Lateral Sclerosis.

Description: The NEDD4 intrachain TR-FRET Assay Kit is a sensitive high-throughput screening (HTS) TR-FRET Assay Kit, designed to measure NEDD4 auto-ubiquitination activity in a homogeneous 384 reaction format. It utilizes a Europium-labeled ubiquitin (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 NEDD4, this assay measures poly-ubiquitination. As a homogeneous assay, it requires no time-consuming washing steps, making it especially suitable for HTS applications as well as real-time analyses.

Product Type: Assay Kit

Shippement Condition: -80°C