Products from BPS Bioscience require a minimum order value above 400€
Application: / Screen molecules that inhibit DCAF5 E3 ligase activity in drug discovery High-Throughput Screening (HTS) applications./ Determine the IC50 of inhibitors of DCAF5 E3 ligase activity./ Determine DCAF5 E3 ligase activity 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. DCAF5 (DDB1 and CUL4 associated factor 15) acts as an E3 ubiquitin ligase when complexed with CUL4A (cullin 4A) or CUL4B. DCAF5 is required for cell survival in SMARCB1-mutant cancers. It acts by increasing the degradation of SWI/SNF (Switch/Sucrose Non-Fermentable) complexes when SMARCB1 (SWI/SNF related, matrix associated, actin dependent regulator of chromatin, subfamily b, member 1) is absent. The removal of DCAF5 in those cases seems to result in a reverse of the cell to a normal status, and thus targeting DCAF5 may become a promising approach for those types of cancer.
Contraindications: / This kit is compatible with up to 1% final DMSO concentration. / Compounds that are fluorescent may interfere with the results, depending on their spectral excitation and emission properties./ It is recommended that the compound alone is tested to determine any potential interference of the compound on the assay results.
Description: The DCAF5 Intrachain TR-FRET Assay Kit is a homogeneous, sensitive TR-FRET (Time-Resolved Fluorescence Resonance Energy Transfer) assay kit designed to measure DCAF5 (DDB1- and CUL4-associated factor 5) complex auto-ubiquitination activity. It utilizes a mix of donor and acceptor labeled ubiquitin 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 DCAF5, making it suitable for real-time kinetics analyses of poly-ubiquitination. The kit contains enough purified DCAF5 complex, purified UBE1 and UbcH5b, detection reagents and assay buffer for 400 reactions.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Target: DCAF5
Uniprot: Q96JK2
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
References: Radko-Juettner S., et al., 2024 Nature 628: 442-449.
Deutsch
English
