Abstract: A method for increasing the light emission produced by the chemiluminescent reaction of 8-amino-5-chloro-7-phenylpyrido[3,4-d]pyridazine-1,4(2H,3H)-dione, a peroxidase enzyme or a conjugate thereof, an enhancer, a co-enhancer and a peroxide oxidizer, wherein the enhancer is an anionic N-alkylphenothiazine and the co-enhancer is selected from a 4-dialkylaminopyridine or an N-azole, and wherein the method comprises the following steps: i. realizing a chemiluminescent substrate by means of mixing together 8-amino-5-chloro-7-phenylpyrido[3,4-d]pyridazine-1,4(2H,3H)-dione, the enhancer, the co-enhancer and the peroxide oxidizer, and ii. adding the peroxidase enzyme or a conjugate thereof to the chemiluminescent substrate.

Abstract: A method for increasing the light emission produced by the chemiluminescent reaction of 8-amino-5-chloro-7-phenylpyrido[3,4-d]pyridazine-1,4(2H,3H)-dione, a peroxidase enzyme or a conjugate thereof, an enhancer, a co-enhancer and a peroxide oxidizer, wherein the enhancer is an anionic N-alkylphenothiazine and the co-enhancer is selected from a 4-dialkylaminopyridine or an N-azole, and wherein the method comprises the following steps: i. realizing a chemiluminescent substrate by means of mixing together 8-amino-5-chloro-7-phenylpyrido[3,4-d]pyridazine-1,4(2H,3H)-dione, the enhancer, the co-enhancer and the peroxide oxidizer, and ii. adding the peroxidase enzyme or a conjugate thereof to the chemiluminescent substrate.

Abstract: A kit for performing an assay for determining an analyte in a sample with an extended shelf-life, wherein the kit comprises a chemiluminescent cyclic dihydrazide, an enhancer, a co-enhancer, and a peroxide oxidizer. The kit is useful in blot assays and immunoassays for the detection of proteins and nucleic acid molecules.

Abstract: A kit for performing an assay for determining an analyte in a sample with an extended shelf-life, wherein the kit comprises a chemiluminescent cyclic dihydrazide, an enhancer, a co-enhancer, and a peroxide oxidizer. The kit is useful in blot assays and immunoassays for the detection of proteins and nucleic acid molecules.

Abstract: A method for increasing the light emission produced by the chemiluminescent reaction of 8-amino-5-chloro-7-phenylpyrido[3,4-d]pyridazine-1,4(2H,3H)-dione, a peroxidase enzyme or a conjugate thereof, an enhancer, a co-enhancer and a peroxide oxidizer, wherein the enhancer is an anionic N-alkylphenothiazine and the co-enhancer is selected from a 4-dialkylaminopyridine or an N-azole, and wherein the method comprises the following steps: i. realizing a chemiluminescent substrate by means of mixing together 8-amino-5-chloro-7-phenylpyrido[3,4-d]pyridazine-1,4(2H,3H)-dione, the enhancer, the co-enhancer and the peroxide oxidizer, and ii. adding the peroxidase enzyme or a conjugate thereof to the chemiluminescent substrate.