OBTENTION OF CYTOPLASM SUB-PROTEOME OF A TROPHOBLAST CELL LINE BY DETERGENT FRACTIONATION
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Novoa-Herrán, S. S., & Sánchez-Gómez, M. (2023). OBTENTION OF CYTOPLASM SUB-PROTEOME OF A TROPHOBLAST CELL LINE BY DETERGENT FRACTIONATION. Revista De La Academia Colombiana De Ciencias Exactas, Físicas Y Naturales, 35(136), 277–286. https://doi.org/10.18257/raccefyn.35(136).2011.2507

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Abstract

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.

https://doi.org/10.18257/raccefyn.35(136).2011.2507

Keywords

Proteomic | cell fractionation | Triton X-100 | digitonine | trophoblast | HTR8/SVneo
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