Claims
- 1. A method for detecting a ligand in a cell or tissue sample, said method comprising,
contacting said sample with a binding agent capable of binding said ligand, wherein said agent is attached to a detectable nucleic acid molecule; staining said sample to identify cells of interest; capturing or isolating said cells of interest; and detecting said nucleic acid molecule wherein the presence of said nucleic acid molecule indicates the presence of said ligand.
- 2. The method of claim 1 wherein said agent is an antibody.
- 3. The method of claim 1 wherein said sample is a tissue section, a cytospin, or a cell smear.
- 4. The method of claim 1 wherein said detecting is by PCR amplification of said nucleic acid molecule.
- 5. The method of claim 4 wherein said PCR is quantitative PCR and the quantitative presence of said ligand is detected.
- 6. The method of claim 1 wherein said staining is by histochemical staining.
- 7. The method of claim 1 wherein said capturing is laser capture microdissection (LCM) or laser microdissection (LMD).
- 8. The method of claim 1 wherein a plurality of agents, attached to a plurality of different nucleic acid molecules, are simultaneously used to detect a plurality of ligands.
- 9. The method of claim 8 wherein said agents are antibodies.
- 10. The method of claim 1 wherein said sample is prostate tissue.
- 11. The method of claim 10 wherein said ligand is prostate specific ligand.
- 12. The method of claim 11 wherein capturing is of only one to two cells.
- 13. The method of claim 1 wherein said nucleic acid molecule comprises a promoter.
- 14. The method of claim 13 wherein said promoter is a T7 promoter.
- 15. The method of claim 14 wherein said detecting comprises contacting said promoter with T7 polymerase and identifying transcription initiated from said T7 promoter.
- 16. The method of claim 8 wherein said plurality of ligands comprise two forms of a polypeptide.
- 17. The method of claim 16 wherein said two forms are the phosphorylated and unphosphorylated forms of a polypeptide.
- 18. The method of claim 5 further comprising quantitatively determining the amount of ligand per captured or isolated cell.
- 19. The method of claim 1 wherein said ligand is a polypeptide, a nucleic acid, a lipid, a carbohydrate, or a portion or domain or epitope thereof.
- 20 The method of claim 15 wherein said identifying is by contacting transcription products with a microarray comprising nucleic acid molecules capable of binding said products by base pair complementarity.
RELATED APPLICATIONS
[0001] This application claims benefit of priority from U.S. Provisional Patent Applications No. 60/271,344, filed Feb. 22, 2001, and 60/314,697, filed Aug. 23, 2001, which are hereby incorporated by reference in their entireties as if fully set forth.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60271344 |
Feb 2001 |
US |
|
60314697 |
Aug 2001 |
US |