Protein probe compounds, compositions, and methods
Coumarine/chinolone and Martina-type compounds, compositions, and method of using the compounds and compositions for protein detection, assay, quantitation, etc. are disclosed. The composition, also referred to as a dye, has enhanced sensitivity over commercially available protein stains, and may be used to stain proteins in solution, proteins separated on gels, proteins transferred to solid supports, etc.
1. A method for staining at least one protein, the method comprising providing a dye comprising compounds V07-05176
and at least one biocompatible excipient to at least one protein to form a dye-protein complex to stain the protein.
2. The method of claim 1 wherein at least one of compound V07-05176 or compound V02-06006 is in the form of a dye-chelate complex.
3. The method of claim 1 wherein a chelating agent of the dye-chelate complex is selected from the group consisting of iminodiacetic acid, nitrilotriacetic acid, 1,2 bis(o-aminophenoxylethane N,N,N′N′ tetraacetic acid, diimines, and combinations thereof, and wherein the chelating agent is chelated to a metal.
4. The method of claim 2 wherein the dye-chelate complex results in increased detergent compatibility.
5. The method of claim 1 wherein the compound further contains a group reactive with at least one of a primary amine, sulfhydryl, carbohydrate, or hydroxyl.
6. The method of claim 5 wherein the group is selected from at least one of hydroxysuccinimide, maleimide, iodoacetamide, hydrazide, or phenyl azide.
7. The method of claim 5 wherein one protein stained is one a protein assay, immunofluorescence assay, singleplex application for protein-protein interactions, or multiplex application for protein-protein interactions.
8. The method of claim 5 wherein one protein stained is one in a multiplex application in combination with other fluorescent dyes or conjugates.
9. A biocompatible composition comprising compounds V02-06006, V07-05176, and at least one biocompatible excipient, where V02-06006 is
and V07-05176 is
10. The method of claim 1 wherein protein detection sensitivity exceeds 3 ng protein.
11. The method of claim 1 wherein protein detection sensitivity exceeds 0.25 ng protein.
12. A biocompatible dye composition comprising at least one excipient and a compound selected from the group consisting of compounds V02-06006, V07-05176, and combinations thereof, in a concentration ranging from about 25 nmol/L to about 500 mmol/L, where compound V02-06006 is
and compound V07-05176 is
13. A method of staining at least one protein, the method comprising
providing a composition comprising at least one excipient and a compound selected from the group consisting of compounds V02-06006, V07-05176, and combinations thereof in an effective concentration to a protein under conditions sufficient for binding the compound to the protein, and
detecting the protein-bound compound,
where compound V02-06006 is
and compound V07-05176 is