Application of iTRAQ multiple labeling and tandem mass spectrometry in proteomic research of human peripheral blood mononuclear cells / 生物医学工程学杂志
Journal of Biomedical Engineering
;
(6): 538-542, 2011.
Artículo
en Chino
| WPRIM
| ID: wpr-359229
ABSTRACT
This paper is aimed to establish and optimize proteomic research platform using isobaric tags for relative and absolute quantitation (iTRAQ) so as to facilitate further proteomic research of human peripheral blood mononuclear cells (hPBMC). We collected hPBMC, after protein extraction and trypsin digestion, we labeled the samples with iTRAQ reagents and then subjected to mass spectrometry. In triplicates, thirty precursors were randomly selected and detected; as a result, 26, 28 and 29 peptides were respectively tagged with ITRAQ reporter ions. The labeling efficiencies ranged between 86.7%-96.7%, with no significant difference among the groups (P>0.05). The coefficient of variance for the relative ratios of peptides from different proteins was ranged from 7.6% to 25.5% and there were no significant differences across the groups (P>0.05). The coefficient of variance for the relative ratios of different peptides from the same protein was varied from 9.3% to 19.1% and the differences across groups were not significant (P>0.05). The labeling of iTRAQ combined with tandem mass spectrometry in PBMC was successful with favourable reproducibility and accuracy, which could lay a foundation for further proteomic study of hPBMC in autoimmune disorders.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Coloración y Etiquetado
/
Leucocitos Mononucleares
/
Proteínas
/
Química
/
Proteoma
/
Proteómica
/
Espectrometría de Masas en Tándem
/
Métodos
Límite:
Humanos
Idioma:
Chino
Revista:
Journal of Biomedical Engineering
Año:
2011
Tipo del documento:
Artículo
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