Intensification of peroxidase-diaminobenzidine staining using gold-sulfide-silver: A rapid and highly sensitive method for visualization in immunoblotting
✍ Scribed by Reiko Iida; Toshihiro Yasuda; Daita Nadano; Koichiro Kishi
- Publisher
- John Wiley and Sons
- Year
- 1990
- Tongue
- English
- Weight
- 435 KB
- Volume
- 11
- Category
- Article
- ISSN
- 0173-0835
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✦ Synopsis
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 α~1~‐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.