1. Reagent preparation
1. SDS-PAGE reagent: see polyacrylamide gel electrophoresis experiment.
2. Homogenization buffer: 1.0 M Tris-HCl (pH 6.8) 1.0 ml; 10% SDS 6.0 ml; β-mercaptoethanol 0.2 ml; ddH2O 2.8 ml.
3. Transfer buffer: Glycine 2.9 g; Tris 5.8 g; SDS 0.37 g; methanol 200 ml; add ddH2O to make up to 1000 ml.
4. 0.01 M PBS (pH7.4): NaCl 8.0 g; KCl 0.2 g; Na2 HPO4 1.44 g; KH2PO4 0.24 g; add ddH2O to 1000 ml.
5. Membrane staining solution: Coomassie brilliant blue 0.2 g; methanol 80 ml; acetic acid 2 ml; ddH2O 118 ml. Coating solution (5% nonfat dry milk, freshly prepared): 1.0 g nonfat dry milk dissolved in 20 ml of 0.01 M PBS.
6. Color developing solution: DAB 6.0 mg; 0.01 M PBS 10.0 ml; nickel sulfate 0.1 ml; H202 1.0 μl.
