Fluorescent dyes and compounds, methods and kits useful for identifying specific organelles and regions in cells of interest
The present invention provides dyes and labeled reagents that may be used in the detection or quantification of desirable target molecules, such as proteins, nucleic acids and cellular organelles. Dyes are provided that may be used free in solution where the binding of the dye to the target molecule provides signal generation. Dyes provided in this invention can comprise reactive groups that may be used to attach the dyes to probes that will bind to desirable target molecules. The novel dyes of the present invention have been substituted with specific groups to provide beneficial properties.
1. A compound having the structure:
wherein X comprises an anion.
2. A compound having the structure:
wherein X comprises an anion.
3. A compound having the structure:
wherein X comprises an anion.
4. A compound having the structure:
wherein X comprises an anion.
5. A compound having the structure:
wherein X comprises an anion.
6. A compound having the structure:
wherein X comprises an anion.
7. A compound having the structure:
wherein X comprises an anion.
8. A compound having the structure:
wherein X comprises an anion.
9. A compound having the structure:
wherein X comprises an anion.
10. A compound having the structure:
wherein X comprises an anion.
11. A compound having the structure:
wherein X comprises an anion and wherein the compound also has at least one reactive group.
12. A compound having the structure:
wherein X comprises an anion.
13. A method of labeling a target molecule comprising the steps of:
(a) providing:
(i) a sample containing said target molecule; and
(ii) the compound of claim 11 ; and
(b) attaching said compound by means of said reactive group to said target molecule in the sample, thereby labeling said target molecule.
14. The compound of claim 11 , wherein said reactive group comprises a nucleophilic reactive group, an electrophilic reactive group, a terminal alkene, a terminal alkyne, a coordinate group or an alkylating agent.
15. The compound of claim 11 , wherein said reactive group is a nucleophilic reactive group that comprises a thiol, amine or hydroxyl group.
16. The compound of claim 11 , wherein said reactive group is an electrophilic reactive group that comprises an isocyanate, isothiocyanate, monochlorotriazine, dichlorotriazine, 4,6,-dichloro-1,3,5-triazines, mono- or di-halogen substituted pyridine, mono- or di-halogen substituted diazine, maleimide, haloacetamide, aziridine, sulfonyl halide, acid halide, hydroxysuccinimide ester, hydroxysulfosuccinimide ester, imido ester, hydrazine, azidonitrophenol, azide, 3-(2-pyridyl dithio)-proprionamide, glyoxal or aldehyde group.
17. A method of identifying a specific organelle or region in a cell of interest comprising the steps of:
(A) providing
(i) said cell of interest; and
(ii) the compound of claim 10 , wherein the compound stains the specific organelle or region;
(B) incubating said cell of interest with said compound; and
(C) identifying the location or said organelle or region in the cell of interest by identifying the compound that stains said organelle or region.
18. The method of claim 17 , wherein said compound is
wherein X comprises an anion, and said organelle or region is endoplasmic reticulum.
19. The method of claim 17 , wherein said compound is
wherein X comprises an anion, and said organelle or region is endoplasmic reticulum.
20. The method of claim 17 , wherein said compound is
wherein X comprises an anion, and said organelle or region is endoplasmic reticulum.
21. The method of claim 17 , wherein said compound is
wherein X comprises an anion, and said organelle or region is a mitochondrion.
22. The method of claim 17 , wherein said compound is
wherein X comprises an anion, and said organelle or region is a mitochondrion.
23. The method of claim 17 , wherein said compound is
wherein X comprises an anion, and said organelle or region is a mitochondrion.
24. The method of claim 17 , wherein said compound is
wherein X comprises an anion, and said organelle or region is a mitochondrion.
25. The method of claim 17 , wherein said compound is
wherein X comprises an anion, and said organelle or region is the whole cell.
26. The method of claim 17 , wherein said compound is
wherein X comprises an anion, and said organelle or region is the whole cell.
27. The method of claim 17 , wherein compound is
wherein X comprises an anion, and said organelle or region is a nucleolus.
28. A kit for identifying an organelle or region in a cell of interest or in a sample containing a cell of interest, said kit comprising in packaged combination:
(A) the compound of claim 10 ;
(B) optional buffers; and
(C) instructions or a protocol for recommended use of the kit.
29. A target molecule comprising the compound of claim 10 .
30. The target molecule of claim 29 , wherein the target molecule is a nucleoside, a nucleotide, an oligonucleotide, a polynucleotide, a peptide nucleic acid, a protein, an oligopeptide, an enzyme, an antibody, a cytokine, avidin, streptavidin, digoxigenin, an oligosaccharide, a polysaccharide, a lipid, a liposome, a glycolipid, or a dye.
31. The target molecule of claim 29 , wherein the target molecule is a protein, an oligopeptide, a nucleotide, an oligonucleotide, or a polynucleotide.
32. The method of claim 13 , wherein the target molecule is a nucleoside, a nucleotide, an oligonucleotide, a polynucleotide, a peptide nucleic acid, a protein, a peptide, an enzyme, an antibody, a cytokine, avidin, streptavidin, digoxigenin, an oligosaccharide, a polysaccharide, a lipid, a liposome, a glycolipid, or a dye.
33. The method of claim 13 , wherein the target molecule is a protein, a peptide, a nucleotide, an oligonucleotide, or a polynucleotide.