Patent ID: 7776533

Claim:
A method for detecting a phosphorylated target molecule in a sample immobilized on a membrane, said method comprising the steps of: i) immobilizing said sample to a membrane; ii) contacting said membrane of step i) with a binding solution, wherein said binding solution comprises: a) phosphate-binding compound having formula (A)m(L)n(B) wherein A is a detectable 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 metal ions; and c) an acid; iii) incubating said membrane of step ii) and said binding solution for sufficient time to allow said phosphate-binding compound to associate with said phosphorylated target molecule; and, iv) detecting said phosphate-binding compound whereby said phosphorylated target molecule is detected, wherein step iii) further comprises contacting said membrane with a wash solution following contact with said binding solution, wherein said phosphorylated target molecule is selected from the group consisting of proteins, peptides, and nucleotides and wherein B of said formula (A)m(L)n(B) comprises a compound selected from the group consisting of Compound 1, 2, 3, 4, 5, 9, 10, 11 and 12 of Table 2.