Protein profiles identified by LC-MS/MS demonstrate change in beta oxidation, ketogenesis, and propionate metabolism in rumen epithelium with different additives

Metalloproteomic analysis of liver proteins isolated from broilers fed with different sources and levels of copper and manganese

“…The protein pellets obtained were solubilized in a specific buffer solution (7 mol L −1 of urea; 2 mol L -1 of thiourea; 2% (w/v) CHAPS (3-[(3-cholaminopropyl)-dimethylammonium]-1-propane sulfonate); 0.5% (v/v) ampholytes pH ranging from 3 to 10; and 0.002% (w/v) bromophenol blue) for performing the liver proteome fractionation process by 2D PAGE (Two-dimensional electrophoresis) according to the procedure already published in previous papers 31 – 36 and briefly described to follow.…”

“…After completing the isoelectric focusing step, the IEF strips were subjected to a hydration process in a reducing solution and later in an alkylating solution 33 , and after this step, the IEF strips were transferred to a 12.5% (v/v) polyacrylamide gel together with a standard containing proteins of different molecular masses and submitted to the separation process in the second dimension (separation according to the molecular mass of the proteins) using the experimental conditions already described in literature 31 – 36 . Finished the 2D PAGE runs, the gels were carefully removed from the glass plates and immersed in a fixative solution containing 10% acetic acid (v/v) and 40% ethanol (v/v) for 30 min.…”