Claims
- 1. A method of assaying for an agent that modulates the attachment of a ubiquitin moiety to at least one ubiquitin agent, said method comprising:
a) combining:
i) a first ubiquitin agent; ii) a candidate agent; and iii) a ubiquitin moiety; and b) assaying for the attachment of said ubiquitin moiety to said first agent.
- 2. The method according to claim 1, wherein said first ubiquitin agent is an ubiquitin activating agent.
- 3. The method according to claim 2, wherein said ubiquitin activating agent is an E1.
- 4. The method according to claim 1, further comprising including a second ubiquitin agent in said combining step.
- 5. The method according to claim 4, wherein said first agent is a ubiquitin conjugating agent and said second agent is a ubiquitin activating agent.
- 6. The method according to claim 5, wherein said ubiquitin conjugating agent is an E2 and said ubiquitin activating agent is an E1.
- 7. The method according to claim 4, wherein said first agent is a ubiquitin ligating agent and said second agent is a ubiquitin conjugating agent comprising said ubiquitin moiety.
- 8. The method according to claim 7, wherein said ubiquitin ligating agent is an E3 and said ubiquitin conjugating agent is an E2 comprising said ubiquitin moiety.
- 9. The method according to claim 5, wherein said ubiquitin conjugating agent is an E2 and said ubiquitin activating agent is an E1 comprising said ubiquitin moiety.
- 10. The method according to claim 9 further comprising including a third ubiquitin agent in said combining step.
- 11. The method according to claim 10, wherein said third agent is a ubiquitin ligating agent.
- 12. The method according to claim 11, wherein said ubiquitin ligating agent is an E3.
- 13. The method according to claim 1, wherein said first ubiquitin agent is attached to a solid support.
- 14. The method according to claims 4, wherein said second ubiquitin agent is attached to a solid support.
- 15. The method according to claim 7, wherein said first ubiquitin agent is attached to a solid support.
- 16. The method according to any of claims 13, 14, and 15, wherein said solid support is a microtiter plate.
- 17. The method according to any of claims 13, 14, and 15, wherein said solid support is a bead.
- 18. The method according to claims 1 or 7, wherein said first agent comprises a tag.
- 19. The method according to claim 4, wherein said second agent comprises a tag.
- 20. The method according to any of claims 1, 4, and 7, wherein said ubiquitin moiety comprises a tag.
- 21. The method according to claims 1 or 7, wherein said first agent comprises an attachment tag.
- 22. The method according to claim 4, wherein said second agent comprises an attachment tag.
- 23. The method according to claims 1 or 7, wherein said first agent comprises a label
- 24. The method according to claim 4, wherein said second agent comprises a label.
- 25. The method according to claims 1 or 7, wherein said first agent comprises an epitope tag.
- 26. The method according to claim 4, wherein said second agent comprises an epitope tag.
- 27. The method according to any of claims 1, 4, and 7, wherein at least a first and a second ubiquitin moiety is used, wherein said first and second ubiquitin moiety moieties comprise different fluorescent labels, and wherein said labels form a fluorescence resonance energy transfer (FRET) pair.
- 28. A method of assaying for an agent that modulates the attachment of a ubiquitin moiety to at least one ubiquitin agent, said method comprising:
a) combining:
i) a first ubiquitin agent comprising a ubiquitin ligating agent; ii) a second ubiquitin agent; iii) a candidate agent; iv) a ubiquitin moiety; and v) a target protein; and b) assaying for the attachment of said ubiquitin moiety to said first agent.
- 29. The method according to claim 28, wherein said second agent is a ubiquitin conjugating agent comprising said ubiquitin moiety.
- 30. The method according to claim 28 further comprising including a third ubiquitin agent in said combining step, wherein said third agent is a ubiquitin activating agent; wherein said substrate and said ubiquitin moiety comprise different fluorescent labels, and wherein said labels form a fluorescence resonance energy transfer (FRET) pair.
- 31. The method according to claim 1 or 28, wherein said ubiquitin moiety is a mammalian ubiquitin moiety.
- 32. The method according to claim 28, wherein said ubiquitin moiety comprises a label.
- 33. The method according to claim 28, wherein said label comprises an epitope tag.
Parent Case Info
[0001] This application is a continuation-in-part application of U.S. patent application Ser. No. 09/826,312, filed Apr. 3, 2001 (pending) which is a continuation-in-part application U.S. patent application Ser. No. 09/542,497, filed Apr. 3, 2000 (pending).
Provisional Applications (1)
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Continuation in Parts (6)
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