Patent ID: 7226740

Claim:
A method for the detection of immobilized poly(amino acids) in a sample, wherein the method comprises: removing, destroying, or dispersing below the critical micelle concentration any biological membranes that are present in said sample; or adding an anionic detergent to the sample, staining solution or combined sample and staining solution mixture; and a) immobilizing said poly(amino acids) on a solid or semi-solid matrix; b) combining said said sample with a staining solution prior, during or after immobilization of said poly(amino acid), wherein said staining solution comprises a compound having formula wherein A represents the atoms necessary to form one to two fused aromatic rings having 6 atoms in each ring, at least one of which is a nitrogen atom, said ring or rings being optionally further substituted one or more times by C 1 –C 6 alkyl, C 1 –C 6 alkoxy trifluoromethyl, halogen, BRIDGE, —L—Rx or —L—Sc; wherein Rx is a reactive group; Sc is a conjugated substance; and L and BRIDGE are independently a single covalent bond, or a covalent linkage; X is O, S, Se, NR 15 , or CR 16 R 17 , where R 15 is H or an alkyl group having 1–6 carbons; and R 16 and R 17 , which may be the same or different, are independently alkyl groups having 1–6 carbons, or R 16 and R 17 taken in combination complete a five or six membered saturated ring; R 2 is selected from the group consisting of —L—Rx, —L—Sc, TAIL, BRIDGE and an alkyl group having 1–6 carbons that is optionally substituted by sulfo, carboxy, amino, substituted amino or substituted ammonium, wherein α is 0 or 1; and TAIL is a heteroatom-containing moiety; Y is —CR 3 ═CR 4 — wherein p and m=0 or 1, such that p+m=1; R 3 , R 4 , R 6 , and R 7 are independently selected from the group consisting of hydrogen, a C 1 –C 6 alkyl, a halogen, a CYCLIC SUBSTITUENT, —OR 8 , —SR 8 , —(NR 8 R 9 ), TAIL; BRIDGE, —L—Rx and —L—Sc; where R 8 and R 9 are independently a C 1 –C 6 alkyl group, 1–2 alicyclic or aromatic rings; or R 8 and R 9 taken in combination are —(CH 2 ) 2 —V—(CH 2 ) 2 — where V is a single bond, —O—, —CH 2 —, or —NR 10 —, where R 10 is H or an alkyl having 1–6 carbons; wherein CYCLIC SUBSTITUENT is a substituted or unsubstituted aryl, heteroaryl or C 3 –C 10 cycloalkyl; or R 6 and R 7 form a fused aromatic ring; R 5 is independently selected from the group consisting of a C 1 –C 6 alkyl group, a CYCLIC SUBSTITUENT, TAIL, BRIDGE, —L—Rx and —L—Sc; or R 5 is absent; R 30 , R 31 , and R 32 are independently selected from the group consisting of hydrogen, C 1 –C 6 alkyl, C 3 –C 10 cycloalkyl, aryl, and heteroaryl, wherein n=0, 1 or 2; wherein, BRIDGE, when present, is bound to a compound having formula I or another unsymmetrical cyanine dye c) incubating said sample and said staining solution for a sufficient amount of time to form a dye-poly(amino acid) complex; d) illuminating said complex with an appropriate wavelength whereby the presence of said poly(amino acid) is determined.