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Título:
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Thiol-free reducing agents in electrophoreticseparations and FASP proteolytic digestions forthe analysis of metal-binding proteins
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Autores:
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Moraleja, I ;
Moreno Gordaliza, Estefanía ;
Mena, María Luz ;
Gómez Gómez, M.Milagros
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Tipo de documento:
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texto impreso
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Editorial:
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Elsevier, 2014
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Dimensiones:
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application/pdf
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Nota general:
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cc_by
info:eu-repo/semantics/openAccess
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Idiomas:
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Palabras clave:
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Estado = Publicado
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Materia = Ciencias: Química: Química analítica
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Tipo = Artículo
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Resumen:
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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.
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En línea:
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https://eprints.ucm.es/id/eprint/55183/1/PDF-on-line%20%28SI%2C%20CC%29.pdf
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