RT Journal Article
T1 Thiol-free reducing agents in electrophoreticseparations and FASP proteolytic digestions forthe analysis of metal-binding proteins
A1 Moraleja, I
A1 Moreno Gordaliza, Estefanía
A1 Mena, María Luz
A1 Gómez Gómez, M.Milagros
AB 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.
PB Elsevier
SN 2215-0161
YR 2014
FD 2014
LK https://hdl.handle.net/20.500.14352/34099
UL https://hdl.handle.net/20.500.14352/34099
LA eng
NO Unión Europea. FP7
NO Ministerio de Ciencia e Innovación (MICINN)
NO Universidad Complutense de Madrid
DS Docta Complutense
RD 27 abr 2024