Labeling of target molecules, identification of organelles and other applications, novel compositions, methods and kits
The present invention provides dyes, reactive 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 are also provided that 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 modified 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 and wherein the compound also has at least one reactive group.
8. 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 7 ; and
(b) attaching said compound by means of said reactive group to said target molecule, thereby labeling said target molecule.
9. The method of claim 8 , 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.
10. The method of claim 9 , wherein said reactive group comprises a nucleophilic reactive group comprising a thiol, amine or hydroxyl group.
11. The method of claim 9 , wherein said reactive group comprises a electrophilic reactive group comprising 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.
12. 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 6 , wherein the compound stains the specific organelle or region;
(B) incubating said cell of interest with said compound; and
(C) identifying the location of said organelle or region in the cell of interest by identifying the compound that stains said organelle or region.
13. The method of claim 12 , wherein the compound is
wherein said organelle or region is a nucleolus.
14. The method of claim 12 , wherein the compound is
wherein said organelle or region is a nucleolus.
15. The method of claim 12 , wherein the compound is
wherein said organelle or region is a nucleolus.
16. The method of claim 12 , wherein the compound is
wherein said organelle or region is a nucleus.
17. The method of claim 12 , wherein the compound is
wherein said organelle or region is a nucleus.
18. A kit for identifying an organelle or a region in a cell of interest, said kit comprising in packaged combination:
(A) the compound of claim 6 or 7 ;
(B) optional buffers; and
(C) instructions or a protocol for recommended use of the kit.
19. The compound of claim 7 , 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.
20. The compound of claim 7 , wherein said reactive group comprises a nucleophilic reactive group comprising a thiol, amine or hydroxyl group.
21. The compound of claim 7 , wherein said reactive group comprises a electrophilic reactive group comprising 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.
22. A target molecule comprising the compound of claim 6 .
23. The target molecule of claim 22 , 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.
24. The target molecule of claim 22 , wherein the target molecule is a protein, an oligopeptide, a nucleotide, an oligonucleotide, or a polynucleotide.
25. The method of claim 8 , 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.
26. The method of claim 8 , wherein the target molecule is a protein, a peptide, a nucleotide, an oligonucleotide, or a polynucleotide.