Objective To compare different staining methods for hippocampal proteomic analysis using 2-dimensional gel electrophoresis (2-DE) and to provide basis for similar researches.
Methods Rat hippocampus were isolated and prepared for total protein samples. Supernatant was determined by Bradford method and separated by 2-DE. Modified Coomassie brilliant blue staining and silver staining were employed for the detection of total protein loaded in samples of different hippocampus protein levels. Specially, Coomassie brilliant blue-silver staining was applied again for the medium loading sample.
Results With the loading protein increasing, the protein spots and impurities were increasing. In the present study, silver staining was ideal for the loading amount of 700 μg protein and modified Coomassie brilliant blue staining for 1 800 μg. Coomassie brilliant blue-silver dual staining was superior to Coomassie brilliant blue staining alone for the 1 200 μg loading sample.
Conclusion More spots can be observed after silver staining which is more sensitive to 2-DE hippocampus separation than modified Coomassie brilliant blue staining. The sensitivity of Coomassie brilliant blue-silver dual staining is better than single modified Coomassie brilliant blue staining.
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