Products from BPS Bioscience require a minimum order value above 400€Applications: Screen molecules that inhibit DCAF11 Ubiquitin ligase activity in drug discovery HTS applicationsDetermine Inhibitor IC
50Perform DCAF11 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 E2ˆ¼Ub conjugate and a substrate protein. This step ensures the transfer of Ub from E2ˆ¼Ub to the substrate, leading to its mono- or poly-ubiquitination.DDB1 and CUL4-associated Factor 11 (DCAF11) is a protein that associates with CUL4A or CUL4B to form a complex with E3 ligase activity. DCAF11 interacts with and ubiquitinates nuclear factor E2-related factor 2 (Nrf2), which is known to regulate cellular stress response genes, including those associated with oxidative stress. Additionally, DCAF11 activity is repressed by transcriptional factor EB (TFEB), allowing for stabilization of proteins such as Nrf2. Because Nrf2 and TFEB are involved in misfolded protein clearance, and oxidative stress is known to contribute to diseases such as Alzheimer's, Huntington's, and Parkinson's disease, DCAF11 is a potential target for the treatment of multiple neurodegenerative disorders. Like most E3 ligases, DCAF11 ubiquitinates itself.
Contraindications: The DCAF11 Intrachain TR-FRET Assay Kit is compatible with up to 1% final DMSO concentration. We recommend preparing the inhibitor in no higher than 5% DMSO solution in buffer and using 4 µl per well.
Description: 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.
Figure 1. DCAF11 Intrachain TR-FRET Assay Kit Schematic.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Uniprot: Q8TEB1
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
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