2026-06-27T01:02:51Zhttps://docta.ucm.es/rest/oai/request

oai:docta.ucm.es:20.500.14352/340992024-10-01T17:59:00Zcom_20.500.14352_14col_20.500.14352_15

Moraleja, I Moreno Gordaliza, María Estefanía Mena Fernández, María Luz Gómez Gómez, María Milagros 2023-06-19T13:35:56Z 2023-06-19T13:35:56Z 2014 2215-0161 10.1016/j.mex.2014.08.003 https://hdl.handle.net/20.500.14352/34099 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4472949/ The analysis of the complexes between metal-based chemotherapeutic drugs and proteins in biological samples, such as cisplatin or oxaliplatin, can be a challenge due to metal strong reactivity towards S-donor molecules such as dithiothreitol (DTT) or b-mercaptoethanol (BME), usually employed as reducing agents in electrophoretic separations and proteolytic digestions for LC–MS/MS analysis. This protocol describes the use of the thiol-free reducing trialkylphosphines, such as tributylphosphine (TBP) and tris(2-carboxyethyl)phosphine (TCEP) as suitable reagents for the preservation of the metal-protein complexes during OFFGEL-IEF and SDS-PAGE separations, respectively. Moreover, the filter-aided sample preparation (FASP) method is presented as an advantageous option to perform tryptic in-solution digestions of metal–protein complexes in combination with OFFGEL-IEF separations. eng https://creativecommons.org/licenses/by/3.0/es/ open access Atribución 3.0 España Thiol-free reducing agents in electrophoretic separations and FASP proteolytic digestions for the analysis of metal-binding proteins journal article