Abstract
We have engineered a monomeric blue non-fluorescent chromoprotein called Ultramarine (fluorescence quantum yield, 0.001; ε(585 nm), 64,000 M(-1) x cm(-1)) for use as a Förster resonance energy transfer acceptor for a number of different donor fluorescent proteins. We show its use for monitoring activation of caspase 3 in live cells using fluorescence lifetime imaging. Ultramarine has the potential to increase the number of cellular parameters that can be imaged simultaneously.
MeSH terms
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Caspase 3 / metabolism
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Chromatography / methods
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DNA Primers / genetics
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Enzyme Activation
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Fluorescence Resonance Energy Transfer / instrumentation*
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Fluorescence Resonance Energy Transfer / methods
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Fluorescent Dyes / chemistry
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Green Fluorescent Proteins / chemistry
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HeLa Cells
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Humans
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Hydrogen-Ion Concentration
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Microscopy / methods
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Mutagenesis, Site-Directed
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Ultracentrifugation / methods
Substances
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DNA Primers
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Fluorescent Dyes
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Green Fluorescent Proteins
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Caspase 3