Bioluminescent metal ion assay
AssigneeRichard B. Thompson
InventorsRichard B. Thompson, Evgenia Matveeva, Carol Fierke, Leslie Bourne, Graham Franke
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Abstract
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.
Claims26
A method for determining metal ions comprising contacting a sample, a cell, or a tissue with a luciferase-conjugated apocarbonic anhydrase;
- contacting the sample, cell or tissue with an ultimate acceptor ligand;
- adding one or more luciferase substrates;
- and measuring light emitted from the sample, cell or tissue.
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The compound LISSAMINE rhodamine B sulfonamide ethyl phenyl sulfonamide.
The compound fluorescein thioureidyl ethyl phenyl sulfonamide.
Dependencies are inferred from the claim text.