OBTENCIóN DE UN SUB-PROTEOMA DE CITOPLASMA DE UNA LíNEA CELULAR DE TROFOBLASTO MEDIANTE FRACCIONAMIENTO CON DETERGENTES

the differential detergent fractionation method described by rambsy and makowski in 1999, was modified using a digitonin (0.015%) buffer to permeate htr8/svneo human trophoblast cells and obtain the cytoplasm proteins in the soluble fraction, while membrane proteins were dissolved from the pellet with triton x-100 (0.5%) buffer. we obtained a reproducible membrane-free cytoplasm protein extract suitable for 2-de, by chloroform/methanol precipitation. in conclusion, the method here described is appropriate for the separation of cytoplasm sub-proteomes from cell cultures, with an optimal reproducibility and recovery to perform proteomic profile analysis by bidimensional electrophoresis.