Chemiluminescent methods and kit

Claim 3

A method according to claim 1, wherein said oxidation catalyst is a peroxidase.

Claim 4

A method according to claim 2, wherein said oxidation catalyst is horseradish peroxidase.

Claim 5

A method according to claim 1, wherein said inorganic peroxide source is hydrogen peroxide.

Claim 7

An assay kit for detecting a component of a chemiluminescent reaction involving a chemiluminescent compound, an inorganic peroxide source and an oxidation catalyst as reactants, said component being one of said chemiluminescent compound, said inorganic peroxide source and said oxidation catalyst, said assay kit comprising: a) said reactants other than said component; and b) an amine selected from the group consisting of: (a) a mono-cyclic aniline compound of the formula: wherein X is hydrogen, methyl, or ethyl; each Y independently is an electron-donating group; and n is zero, one, two or three; (b) a biphenyl compound of the formula: wherein X is hydrogen, methyl or ethyl; W is a carbon-carbon bond, oxygen, methylene, or -CH₂O-; and Z and Z′ are independently hydrogen, halogen, amino, lower alkyl or lower alkoxy; (c) a naphthalene compound of the formula: wherein X is hydrogen, methyl or ethyl; and Z and Z′ are independently hydrogen, halogen, amino, lower alkyl or lower alkoxy; and (d) an anthracene compound of the formula: wherein X is hydrogen, methyl or ethyl; and Z and Z′ are independently hydrogen, halogen, amino, lower alkyl or lower alkoxy; said amine being further characterized by the fact that it provides a chemiluminescent light output signal which, one minute after peak light output is attained, is at least about 50% of the intensity of said peak light output when the chemiluminescent reaction is conducted in a reaction mixture obtained by combining i) 100 microliters of an aqueous solution containing luminol at a concentration of 0.01 mg/ml and 0.05 M sodium borate/0.01 M EDTA buffer to maintain the pH at 9.5; ii) 10 microliters of a 0.04 mg/ml aqueous solution of horseradish peroxidase; iii) 100 microliters of a 0.035 M solution of hydrogen peroxide in 0.001 M EDTA; iv) 100 microliters of water; and v) 10 microliters of one and/or the other of a 0.1 mg/ml or a 1 mg/ml aqueous solution of the amine.

Claim 8

An assay kit for detecting a component of a chemiluminescent reaction involving a chemiluminescent compound, an inorganic peroxide source and an oxidation catalyst as reactants, said component being one of said chemiluminescent compound, said inorganic peroxide source and said oxidation catalyst, said assay kit comprising: a) said reactants other than said component; and b) an amine selected from the group consisting of: (a) a mono-cyclic aniline compound of the formula: wherein X is hydrogen, methyl, or ethyl; each Y independently is an electron-donating group; and n is zero, one, two or three; (b) a biphenyl compound of the formula: wherein X is hydrogen, methyl or ethyl; W is a carbon-carbon bond, oxygen, methylene, or -CH₂O-; and Z and Z′ are independently hydrogen, halogen, amino, lower alkyl or lower alkoxy; (c) a naphthalene compound of the formula: wherein X is hydrogen, methyl or ethyl; and Z and Z′ are independently hydrogen, halogen, amino, lower alkyl or lower alkoxy; and (d) an anthracene compound of the formula: wherein X is hydrogen, methyl or ethyl; and Z and Z′ are independently hydrogen, halogen, amino, lower alkyl or lower alkoxy; said amine being further characterized by the fact that it provides a chemiluminescent light output signal which, one minute after peak light output is attained, is at least about 50% of the intensity of said peak light output when the chemiluminescent reaction is conducted in a reaction mixture obtained by combining i) 100 microliters of an aqueous solution containing luminol at a concentration of 0.01 mg/ml and 0.05 M sodium borate/0.01 M EDTA buffer to maintain the pH at 9.5; ii) 10 microliters of a 0.04 mg/ml aqueous solution of horseradish peroxidase; iii) 100 microliters of a 0.035 M solution of hydrogen peroxide in 0.001 M EDTA; iv) 100 microliters of water; and v) 10 microliters of one and/or the other of a 0.1 mg/ml or a 1 mg/ml aqueous solution of the amine.