Assay methods and systems
View Patent ↗Methods, systems, kits for carrying out a wide variety of different assays that comprise providing a first reagent mixture which comprises a first reagent having a fluorescent label. A second reagent is introduced into the first reagent mixture to produce a second reagent mixture, where the second reagent reacts with the first reagent to produce a fluorescently labeled product having a substantially different charge than the first reagent. A polyion is introduced into at least one of the first and second reagent mixtures, and the fluorescent polarization in the second reagent mixture relative to the first reagent mixture is determined, this fluorescent polarization being indicative of the rate or extent of the reaction.
1. A kit, comprising:
a fluorescently labeled phosphorylatable compound which is capable of being phosphorylated to produce a fluorescently labeled phosphorylated product; and
a liquid reagent comprising a polymer having multivalent metal cations associated therewith, wherein the multivalent metal cations bind the polymer to the phosphrylated product.
2. The kit of claim 1 , wherein the multivalent metal cations comprise trivalent metal cations.
3. The kit of claim 1 , wherein the multivalent metal cations are selected from a group consisting of Fe3+, Ca2+, Ni2+ and Zn2+.
4. The kit of claim 1 , wherein the phosphorylatable compound is selected from a group consisting of a serine, threonine or tyrosine substrate.
5. The kit of claim 1 , wherein the multivalent metal cations are chelated to the polymer.
6. The kit of claim 1 , wherein the polymer is between 5 kD and 1000 kD.
7. The kit of claim 1 , further comprising a kinase enzyme.
8. The kit of claim 1 , further comprising ATP.
9. The kit of claim 1 , further comprising a buffer solution.
10. The kit of claim 1 , wherein the multivalent metal cations comprise Fe3+.
11. The kit of claim 1 , further comprising a multiwell plate.
12. A kit, comprising:
a fluorescently labeled phosphorylatable compound which is capable of being phosphorylated to produce a fluorescently labeled phosphorylated product; and
a liquid reagent comprising a molecule having multivalent metal cations associated therewith, wherein the metal cations bind the molecule to the phosphorylated product.
13. The kit of claim 12 , wherein the multivalent metal cations comprise trivalent metal cations.
14. The kit of claim 12 , wherein the multivalent metal cations are selected from a group consisting of Fe 3+, Ca2+, Ni2+ and Zn2+.
15. The kit of claim 12 , wherein the phosphorylatable compound is selected from a group consisting of a serine, threonine or tyrosine substrate.
16. The kit of claim 12 , wherein the molecule comprises a polymer.
17. The kit of claim 16 , wherein the multivalent metal cations are chelated to the polymer.
18. The kit of claim 17 , wherein the polymer is between 5 kD and 1000 kD.
19. The kit of claim 12 , further comprising a kinase enzyme.
20. The kit of claim 12 , further comprising ATP.
21. The kit of claim 12 , further comprising a buffer solution.
22. The kit of claim 12 , further comprising a multiwell plate.