b'Example data12345678910Figure 35. Sensitive staining results with SimplyBlue SafeStain. TheFigure 36. Two-dimensional electrophoresis analysis of spinach following samples were separated on an Invitrogen NuPAGE 412%chloroplast extract; staining with SimplyBlue SafeStain. A spinach Bis-Tris gel and then stained with SimplyBlue SafeStain. Lane 1: 6 gchloroplast extract was prefractionated in the Invitrogen ZOOM IEF protein mix; lane 2: 1 g rabbit IgG; lane 3: 1 g reduced BSA; lane 4:Fractionator, and the individual fractions were then separated by 2D 5 g E. coli lysate; lane 5: 20 ng reduced BSA; lane 6: 10 ng reducedelectrophoresis using narrow pHrange Invitrogen ZOOM IPG strips and BSA; lane 7: 7 ng reduced BSA; lane 8: 3 ng reduced BSA; lane 9: 10 Lan Invitrogen NuPAGE 412% Bis-Tris ZOOM gel. Gels were stained Invitrogen Mark12 Unstained Standard (blend of 12 purified proteins);using SimplyBlue SafeStain.lane 10: 5 L Mark12 Unstained Standard.1 2 3 4 5 6 7 8 9 1 2 3 4 5 6 7 8 9 1 2 3 4 5 6 7 8 9Rabbit IgGBSAProtein AProtein GLysozyme5-minute stain; 1-hour stain; 1-hour stain;15-minute water destain 2-hour water destain overnight water destainFigure 37. Enhanced sensitivity and clear background using Imperial Protein Stain. For even greater sensitivity and reduced background, gels can be stained with Imperial Protein Stain for 1 hour and washed with water from 1 hour to overnight. Lane 1: BSA only (6 g); lanes 29: loaded with 1,000, 200, 100, 50, 25, 12, 6, and 3 ng protein.Did you knowStaining with a Coomassie stain prior to silver staining allows for more uniform staining of certain proteins since silver ions can interact with certain functional groups such as carboxylic acid groups, imidazole, sulfhydryls, and amines.66'