Intensification of peroxidase-diaminobenzidine staining using gold-sulfide-silver: a rapid and highly sensitive method for visualization in immunoblotting. |
| |
Authors: | R Iida T Yasuda D Nadano K Kishi |
| |
Institution: | Department of Legal Medicine, Fukui Medical School, Japan. |
| |
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 alpha 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. |
| |
Keywords: | |
|
|