1129. Ad.Interferon Beta Is an Effective Treatment in an Orthotopic Model of Broncho-Alveolar Non-Small Cell Lung Cancer Via NK Cell and CD 8+ T Cell Mechanisms

Wilderman, Michael; Sun, Junsong; Vachani, Anil; Suzuki, Eiji; Sterman, Dan H.; Kaiser, Larry R.; Albelda, Steve M.

doi:10.1016/j.ymthe.2005.07.677

Cited by 1 publication

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“…Gluc is a smaller and brighter luciferase known to date and was cloned from a marine copepod (Tannous et al, 2005; Welsh et al, 2009). It shares some spectral properties with Rluc and has been recently used in BRET1 assays (Li et al, 2012).…”

Section: Which Bret Version To Chose?mentioning

confidence: 99%

Setting Up a Bioluminescence Resonance Energy Transfer High throughput Screening Assay to Search for Protein/Protein Interaction Inhibitors in Mammalian Cells

Couturier¹,

Deprez²

2012

Front. Endocrin.

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Each step of the cell life and its response or adaptation to its environment are mediated by a network of protein/protein interactions termed “interactome.” Our knowledge of this network keeps growing due to the development of sensitive techniques devoted to study these interactions. The bioluminescence resonance energy transfer (BRET) technique was primarily developed to allow the dynamic monitoring of protein/protein interactions (PPI) in living cells, and has widely been used to study receptor activation by intra- or extra-molecular conformational changes within receptors and activated complexes in mammal cells. Some interactions are described as crucial in human pathological processes, and a new class of drugs targeting them has recently emerged. The BRET method is well suited to identify inhibitors of PPI and here is described why and how to set up and optimize a high throughput screening assay based on BRET to search for such inhibitory compounds. The different parameters to take into account when developing such BRET assays in mammal cells are reviewed to give general guidelines: considerations on the targeted interaction, choice of BRET version, inducibility of the interaction, kinetic of the monitored interaction, and of the BRET reading, influence of substrate concentration, number of cells and medium composition used on the Z′ factor, and expected interferences from colored or fluorescent compounds.

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Section: Which Bret Version To Chose?mentioning

confidence: 99%