XIAP Intrachain TR-FRET Assay Kit

Products from BPS Bioscience require a minimum order value above 400€

Applications: Great for screening molecules that inhibit XIAP Ub ligase activity HTS applications in drug discovery, for determination of compound IC₅₀ and for XIAP real-time kinetics analyses.

Contraindications: The XIAP 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: 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.The X-linked inhibitor of apoptosis (XIAP) protein is a RING-containing E3 Ub ligase which has the ability to directly regulate caspases and suppress apoptotic cell death pathways. An increased expression level of XIAP has been shown for many cancer types and is associated with cancer cell migration. Like most E3 ligases, XIAP ubiquitinates itself.The XIAP intrachain TR-FRET Assay Kit is a sensitive high-throughput screening (HTS) TR-FRET Assay Kit, designed to measure XIAP auto-ubiquitination activity in a homogeneous 384 reaction format. It utilizes a Europium cryptate-labeled 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 XIAP, this FRET-based assay requires no time-consuming washing steps, making it especially suitable for HTS applications as well as real-time kinetics analyses of polyubiquitination.

Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.

Supplied As: Homogeneous 384 reaction format

Warnings: Avoid freeze/thaw cycles.

Biosafety Level: Not applicable (BSL-1)

• SKU: BPS78306
• Manufacturer: BPS Bioscience
• Manufacturer SKU: 78306
• Green Labware: No
• Package Unit: 384 rxns.
• Quantity Unit: PAK