Other

Carol Ann Fierke

Richard B Thompson

Leslie Bourne

Michele Cramer

Hui Hui Zeng

Evgenia Matveeva

Graham Franke

Rebecca Bozym

A method for determining metal ions, both qualitatively and quantitatively, is disclosed. The method utilizes emission from fluorescence resonance energy transfer from a luciferase-carbonic anhydrase conjugate or fusion protein to an acceptor ligand in the presence of metal ion bound to the protein to measure free metal ion concentrations down to picomolar concentration ranges. The method is relatively insensitive to contaminants, and so can be used to measure metal ion concentrations in cells, body fluids or environmental samples without extensive sample preparation.

Bioluminescent metal ion assay

A highly selective and sensitive of a carbonic anhydrase-based method for measurement of zinc ion by an excitation ratiometric format based on resonance energy transfer: i.e., where the zinc ion level is transduced as the ratio of fluorescence intensities excited at two different excitation wavelengths, is provided. The method can be used very well in a fluorescence microscopy format. A detection limit of about 10 pM in zinc buffered systems, a ten to one thousand-fold improvement on the Fura indicators (which respond to Ca and Mg as well), and a one hundred thousand-fold improvement on the recently described FuraZin-1 is achieved.

Excitation ratiometric fluoroscent biosensor for zinc ion at picomolar levels

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Ann Harbor, MI, United States of America

Carol Ann Fierke