title: Intensification of peroxidase-diaminobenzidine staining using gold-sulfide-silver: a rapid and highly sensitive method for visualization in immunoblotting.
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.
authors.affiliation: Department of Legal Medicine, Fukui Medical School, Japan.
authors.forename: R
authors.initials: R
authors.lastname: Iida
authors.forename: T
authors.initials: T
authors.lastname: Yasuda
authors.forename: D
authors.initials: D
authors.lastname: Nadano
authors.forename: K
authors.initials: K
authors.lastname: Kishi
mesh_terms: D003850:Deoxyribonuclease I / Q000235:genetics / Q000652:urine; D004591:Electrophoresis, Polyacrylamide Gel; D006046:Gold; D015151:Immunoblotting / Q000379:methods*; D007525:Isoelectric Focusing; D010544:Peroxidases; D010641:Phenotype; D012680:Sensitivity and Specificity; D012834:Silver; D013194:Staining and Labeling; D013440:Sulfides; D014168:Transferrin / Q000235:genetics; D014809:Vitamin D-Binding Protein / Q000235:genetics; D000515:alpha 1-Antitrypsin / Q000235:genetics
chemical_list: D013440:Sulfides; D014168:Transferrin; D014809:Vitamin D-Binding Protein; D000515:alpha 1-Antitrypsin; D012834:Silver; D006046:Gold; D010544:Peroxidases; C037561:diaminobenzidine peroxidase; D003850:Deoxyribonuclease I