For detection of proteins and nucleic acids migration by polyacrylamide gel electrophoresis, silver staining is now being highlighted as a sensitive staining method. However the original silver staining method is not necessarily easy to work and takes long-time. 2D-SILVER STAIN-II is an improved reagent kit developed to provide simple and fast staining.
2D-SILVER STAIN? is applicable to the detection of proteins and nucleic acids subjected to polyacrylamide and SDS-polyacrylamide slab gel electrophoresis.
Running Condition
・Loading buffer： Tris SDS
・Sample： PageRuler™ Prestained Protein Ladder #SM1811（Fermentas UAB）
| ? ×4 | ? ×128 |
| ? ×8 | ? ×256 |
| ? ×16 | ? ×512 |
| ? ×32 | ? ×1024 |
| ? ×64 | |
| (Each Lane 3) | |
・Electrode buffer： SDS-Tris-Glycine Electrode buffer
・Running time：30 mA constant current/sheet、About 50 min.
・Dying Method: 2D-Silver Stain
1. Rapid staining results in a short time after electrophoresis.
2. Staining with higher sensitivity For proteins:50 to 100 times more sensitive than CBB staining For nucleic acids: 50 to 100 times more sensitive than EB staining
3. Simple preparation and simple operation.
4. No use of such materials as heavy metals controlled by regulations.
5. Capable of restaining CBB-stained gels. (Double-staining)
6. Staining process can be stopped any time during development for desired chromatic figures.
| Regents | Main ingredients | Volume |
|---|---|---|
| 1) Fixing Reagent | Thiourea | 100mLx1 |
| 2) Pretreatment reagent | Dithiothreitol, Glutar-aldehyde, Thiourea | 100mLx1 |
| 3) Staining Solution A | Siver nitrate | 100mLx1 |
| 4) Staining Solution B | Ammonium hydroxide, Sodium hydroxide | 100mLx1 |
| 5) Concentrated developer | Citric acid, Folmaldehyde, Sodium hydroxide | 100mLx1 |
| 6) Stopper | Citric acid | 100mLx1 |
<Reference>
1. K. Ohsawa et. Al.: Silver Stain for Detecting 10-Femotogram Quantities of protein after Polyacrylamide Gel Electrophoresis, Anal. Biochem., 135, 409(1983)
2. S. Irie: A highly sensitive silver Staining for detection of proteins in polyacrylamide gels, biochemistry(Japan), 52, 411(1980)
3. B.R. Oakley et. al.: Visualization of proteins with silver “stain” for detecting proteins in polyacrylamide gels, Anal. Biochem., 105, 361(1980)
4. H.M. Poehling et. al.: Visualization of proteins with silver ”stain”, a critical analysis, electrophoresis, 2, 141 (1981)
5. U.K. Laemmli: Cleavage of structural proteins during the assembly of the head of bacteriophage T4., Nature, 227, 680(1970)
