Chemical signal enhancement of dynamic intensity-based...

G - Physics – 01 – N

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G01N 33/50 (2006.01) G01N 33/58 (2006.01)

Patent

CA 2407521

The present invention involves that adding certain dyes to the solution outside cells expressing a GFP-tagged proteins in a redistribution assay, enhance the signal. Examples of such dyes are Trypan Blue and Acid Red. This enables redistribution assays to be measured as a change in light intensity. Redistribution movement from the cytosol to the membrane, or vice versa, are illustrated. The apparatus used for measuring the redistribution is an ordinary plate reader or a FLIPRTM type instrument. It is a common feature among the dyes described that the enhancer compound has an absorption spectrum overlapping the emission spectrum of the GFP.

La présente invention constate que l'ajout de certains colorants à la solution extérieure des cellules exprimant une protéine marquée GFP dans un dosage de redistribution, renforce le signal. Parmi ces colorants, on trouve le bleu trypan et le rouge acide. Ils permettent de mesurer des dosages de redistribution sous forme de modification à l'intensité lumineuse. On présente le mouvement de redistribution en provenance du cytosol en direction de la membrane, ou vice-versa. L'appareil utilisé pour mesurer cette redistribution est un lecteur de plaque ordinaire ou un instrument de type FLIPRTM (lecteur de plaque par imagerie fluorométrique). Ces colorants possèdent une caractéristique commune en ce sens que le composé de renfort présente un spectre d'absorption chevauchant l'émission spectrale de la protéine GFP.

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