Compositions and methods for detection and isolation of phosphorylated molecules
First Claim
1. A binding solution comprising:
- a) a BAPTA metal chelating moiety;
b) a salt comprising trivalent metal ions; and
,c) an acid.
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Abstract
The present invention relates to phosphate-binding compounds that find use in binding, detecting and isolating phosphorylated target molecules including the subsequent identification of target molecules that interact with phosphorylated target molecules or molecules capable of being phosphorylated. A binding solution is provide that comprises a phosphate-binding compound, an acid and a metal ion wherein the metal ion simultaneously interacts with an exposed phosphate group on a target molecule and the metal chelating moiety of the phosphate-binding compound forming a bridge between the phosphate-binding compound and a phosphorylated target molecule resulting in a ternary complex. The binding solution of the present invention finds use in binding and detecting immobilized and solubilized phosphorylated target molecules, isolation of phosphorylated target molecules from a complex mixture and aiding in proteomic analysis wherein kinase and phosphatase substrates and enzymes can be identified.
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Citations
25 Claims
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1. A binding solution comprising:
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a) a BAPTA metal chelating moiety; b) a salt comprising trivalent metal ions; and
,c) an acid. - View Dependent Claims (20, 21, 22, 23, 24, 25)
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2. A binding solution comprising:
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a) a phosphate-binding compound having formula (A)m(L)n(B) wherein A is a chemical moiety, L is a linker, B is a metal-chelating moiety, m is an integer from 1 to 4 and n is an integer from 0 to 4; b) a salt comprising trivalent metal ions; and
,c) an acid. - View Dependent Claims (3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
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Specification