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