Abstract
Relationship Between the Types of the Extraction Buffer with Quality of the 2-Dimensional Electrophoresis Proteomic Map
Highlights
AnimalsOne fourteen-weeks old male Sprague Dawley rat was selected
The percentage of volume and intensity of the detected spots in the Tris-hydrochloric acid (HCl) and rehydration buffers were higher than PBS buffer, but, there were no significant differences neither in %volume, nor in %intensity between three groups
Hydrochloric acid (HCl) and sodium hydroxide were purchased from Merk
Summary
One fourteen-weeks old male Sprague Dawley rat was selected. Left testes frozen and stored in -70 °C until sodium dodecylsulfate–polyacrylamide gel electrophoresis (SDSPAGE) and 2DE gel analysis. Appropriate sample preparation is critical for obtaining high quality two-dimensional electrophoresis (2-DE) separations and reliable results in a proteomic analysis [1,2]. Sample solution must be prepared by using appropriate buffer. The 2-DE sample rehydration buffer (RHB) is widely used to denature and solubilize protein samples. Phosphate saline (PBS) and Trishydrochloride (Tris-HCl) buffers are used for sample preparation for 2-DE and other analytical methods. We compare 2-DE gel patterns obtained by rat testes tissue preparation using the three conventional distinct buffers, Tris-HCl, PBS and RHB
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