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Visualization of Antigen on Nitrocellulose Membrane by the Oxidative Coupling Reaction of N,N'-dimethyl-p-phenylenediamine and 4-chloro-1-naphthol

Visualization of Antigen on Nitrocellulose Membrane by the Oxidative Coupling Reaction of N,N'-dimethyl-p-phenylenediamine and 4-chloro-1-naphthol

R Kobayashi, Y Tashima

Anal Biochem. 1989 Nov 15;183(1):9-12.

PMID: 2482680

Abstract:

A method which allows the highly sensitive and simple immunodetection of antigen-antibody complexes on nitrocellulose papers has been developed. The method is a modification of the procedure known as the "Nadi reaction" (oxidative coupling of 1-naphthol and N,N'-dimethyl-p-phenylendiamine) for histochemical purposes. Proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrophoretically transferred to nitrocellulose membranes. Bound proteins were first reacted with a primary antibody and then with a horseradish peroxidase-labeled second antibody. The antigen-antibody complexes on the membranes were visualized with the oxidative coupling solution containing N,N'-dimethyl-p-phenylendiamine and 4-chloro-1-naphthol. Sensitivity was enhanced 4 to 16 times by the new method relative to that of the 4-chloro-1-naphthol method or the 3,3'-diaminobenzidine method.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42411223	4-Chloro-1-naphthol solution			Price

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