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Sequential or simultaneous emission detection for BRET assays

Marjan Orban BMG LABTECH 12/2014

Bioluminescence Resonance Energy Transfer (BRET) is a system of choice for monitoring intermolecular interactions such as proteomics applications, receptor research and signal transduction. The assay is based on non-radiative energy transfer between fusion proteins containing Renilla luciferase (Rluc) and e.g. Yellow Fluorescent Protein (YFP).

 

The BRET2™ demo kit has been used to prove the feasibility of performing a BRET assay on a BMG LABTECH microplate reader. The BRET demo kit applies the cell-permeable and nontoxic substrate DeepBlueC™ (DBC) and a mutant of the Green Fluorescent Protein (GFP2) as acceptor. These compounds show improved spectral resolution and sensitivity over earlier variants.

 

Ratiometrically quantifiable BRET2™ assays have been successfully measured using either sequential or simultaneous dual emission. However, the simultaneous dual emission option will lead to a higher assay window and is a lot faster. The internal reagent injectors for 384-well plate format offer a great advantage for this kind of assay.

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