Abstract

NOVOA-HERRAN, Sandra Susana  and  SANCHEZ-GOMEZ, Myriam. OBTENCIÓN DE UN SUB-PROTEOMA DE CITOPLASMA DE UNA LÍNEA CELULAR DE TROFOBLASTO MEDIANTE FRACCIONAMIENTO CON DETERGENTES. Rev. acad. colomb. cienc. exact. fis. nat. [online]. 2011, vol.35, n.136, pp.277-285. ISSN 0370-3908.

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.

Keywords : Proteomic; cell fractionation; Triton X-100; digitonine; trophoblast; HTR8/SVneo.

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