TR-FRET In-Vitro Ubiquitylation Assay

* Please be kindly noted that our services and products can only be used for research to organizations or companies and not intended for any clinical or individuals.

Time-resolved fluorescence resonance energy transfer (TR-FRET) is a technology that can be used for in vitro ubiquitylation testing. BOC Sciences has a mature TR-FRET technology platform, and also has a technical team with many years of experience in this technology. As a customer of BOC Sciences, you can discuss research goals and measurement parameters with our technical team. We will customize a suitable cooperation plan based on the discussion results, and provide you with detailed experimental data and analysis reports in a relatively short time.

Introduction

TR-FRET technology combines the advantages of time-resolved fluorescence (TRF) and fluorescence resonance energy transfer (FRET) technologies. FRET technology utilizes the energy transfer between the donor fluorophore and the acceptor fluorophore combined by two biomolecules to emit light at a specific wavelength. However, in the traditional FRET technology, the background fluorescence signal is easily affected by the sample components, such as buffers, proteins, compounds, and cell lysates, which affect the experimental sensitivity and increases the complexity of data analysis. TRF technology can utilize the very long emission half-life of the lanthanides in the rare earth elements, which can remove the background with a short half-life, thereby reducing analytical interference and improving data quality. TR-FRET technology can be used to monitor the formation of polyubiquitin chains by analyzing the ubiquitin combined with the lanthanide fluorophore and the receptor fluorophore to perform in-vitro ubiquitylation assay. Moreover, fluorophore-labeled ubiquitin combined with terbium-anti-epitope-tagged antibody can also be utilized for mono-ubiquitylation detection.

Our Services for TR-FRET In-Vitro Ubiquitylation Assay (include but not limited to the following):

• Ubiquitin Conjugating Assay
  TR-FRET in-vitro ubiquitylation assay not only can be utilized to detect mono-ubiquitylation and also can be used to detect poly-ubiquitylation. It is often used for the detection of E3 ubiquitin ligase, such as mouse double minute 2 homolog (MDM2), neural precursor cell expressed developmentally down-regulated protein 4 (NEDD4) and X-linked inhibitor of apoptosis protein (XIAP), monitoring the binding of ubiquitin to E3 ubiquitin ligase in the enzyme solution. In addition, it can be used for the detection of noncanonical ubiquitin conjugating enzyme (E2) and E2 conjugated ubiquitin protein. Our TR-FRET technology platform can help customers realize the monitoring of ubiquitin binding.
• Ubiquitin Deconjugation Assay
  Uncoupling enzymes can specifically act on the ubiquitin or ubiquitin-like protein (UBL) C-terminal peptide or isopeptide bond, so that the target protein will not be degraded by proteases. TR-FRET technology can be used to measurement the decoupling of ubiquitin using deubiquitinating enzymes (DUBs). The experts of BOC Sciences can work out the needs of customers to formulate suitable ubiquitin deconjugation detection schemes.

Our Advantages

• Advanced TR-FRET technology platform and experienced expert team can develop high-quality test solutions for customers
• Homogeneous operation, no time-consuming washing steps and separation steps
• Low background and little interference from compounds and buffers
• High sensitivity and efficient detection
• Suitable for high-throughput screening
• Data analysis, detailed report with results and discussion
• Quality one-stop service

Workflow of TR-FRET In-Vitro Ubiquitylation Assay

Reference:

1. Madiraju, C.; et al. TR-FRET-based high-throughput screening assay for identification of UBC13 inhibitors. Journal of Biomolecular Screening. 2012, 17(2): 163-176.

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