Intensification of peroxidase‐diaminobenzidine staining using gold‐sulfide‐silver: A rapid and highly sensitive method for visualization in immunoblotting

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Abstract
A highly sensitive and rapid visualization method for protein detection by immunoblotting is described. Proteins blotted onto a Durapore membrane were visualized by the following procedure: after conventional peroxidase‐based staining with 3,3′‐diaminobenzidine (DAB), the produced DAB precipitates were intensified by treating with (i) gold trichloride (acid), (ii) sodium sulfide, and (iii) a developer containing silver nitrate. This postintensification method was employed for the detection of the genetic polymorphism of human proteins, such as deoxyribonuclease I in urine, and group specific component, transferrin and α1‐antitrypsin in serum after polyacrylamide gel‐isoelectric focusing, followed by immunoblotting. This postintensification technique was found to be simple, giving up to 16‐ to 64‐fold amplification of the conventional peroxidase‐DAB staining.

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