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 II is applicable to the detection of proteins and nucleic acids subjected to polyacrylamide and SDS-polyacrylamide slab gel electrophoresis.
• Loading buffer: Tris SDS
• Sample: PageRuler™ Prestained Protein Ladder #SM1811(Fermentas UAB)
|(1) x4||(6) x128|
|(2) x8||(7) x256|
|(3) x16||(8) x512|
|(4) x32||(9) x1024|
|(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.
|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|
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)
2D-Silver Stain Reagent II [For 10 gels ]