Silver stain for detecting 10-femtogram quantities of protein after polyacrylamide gel electrophoresis
β Scribed by Kazuaki Ohsawa; Nobuyosi Ebata
- Publisher
- Elsevier Science
- Year
- 1983
- Tongue
- English
- Weight
- 526 KB
- Volume
- 135
- Category
- Article
- ISSN
- 0003-2697
No coin nor oath required. For personal study only.
β¦ Synopsis
A rapid and highly sensitive silver stain and color stain were developed for visualizing proteins. The procedure is simple and the bands were clear. This silver stain detects 100 pg quantities of proteins. In order to stain quickly, sensitively, and sharply a protein matrix in a gel, the repeated shrinkage and swelling gel was developed with a hyper- and hypotonic solution to remove the sodium dodecyl sulfate (SDS) from SDS-protein complex and to generate influx of staining solution into the gel. We have found that the silver staining method with the repeated exposure to hyper- and hypotonic solution and a narrow well produced 10 fg order of proteins.
π SIMILAR VOLUMES
We have simplified the highly sensitive silver stain of R. C. Switzer III, C. R. Merril, and S. Shifrin (1979, Anal. Biochem. 98, 231β237) for visualizing proteins in polyacrylamide gels. We have reduced the number of steps in the procedure from 10 to 6, simplified the reagents in each step, and red
Methods for the detection of RNase after polyacrylamide gel electrophoresis have required long incubation times or indirect techniques that allowed the enzyme ample time to diffuse into a broad band (l-3). Wolf (4) recently described a new technique in which the gel was incubated in a solution of lo
## Standardization of protein position in silver-stained two-dimensional poly acrylamide gel electrophoresis Methods for standardization of silver-"stained" protein position in two-dimensional polyacrylamide gel electrophoresis are presented for their application to simultaneous multiple gel syste
A fast and convenient method for silver staining of proteins on electroblotting membranes was developed based on Gallyas' histochemical intensifier and applied to human endothelial cell proteins separated by one-and two-dimensional electrophoresis and electroblotted to polyvinyl difluoride membranes