Moraleja, IMoreno Gordaliza, María EstefaníaMena Fernández, María LuzGómez Gómez, María Milagros2023-06-192023-06-1920142215-016110.1016/j.mex.2014.08.003https://hdl.handle.net/20.500.14352/34099The 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.engAtribución 3.0 Españahttps://creativecommons.org/licenses/by/3.0/es/journal articlehttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4472949/open access543Metal-binding proteinsOFFGEL-IEFFASPICP-MSThiol-free reductantsnLC–ESI-LTQ-MS/MSQuímica analítica (Química)2301 Química Analítica