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Enhanced reproducibility of the human gel‐based tear proteome maps in the presence of di‐(2‐hydroxyethyl) disulfide
Author(s) -
SaraygordAfshari Neda,
NaderiManesh Hossein,
Naderi Mostafa
Publication year - 2014
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.1221
Subject(s) - dithiothreitol , reproducibility , proteome , disulfide bond , chromatography , chemistry , polyacrylamide gel electrophoresis , gel electrophoresis , biochemistry , enzyme
Patterns obtained in two‐dimensional gel electrophoresis (2‐ DE ) in the previously published articles suggest a varying number of proteins. To seek the cause of this variation, we investigated the effect of reduction power on the overall tear proteome maps. To this end, the buffers of two reducing agents, dithiothreitol ( DTT ) at nine different concentrations and di‐(2‐hydroxyethyl) disulfide ( HED ), were examined. The assay showed that HED clearly improved 2‐ DE resolution, increased the number of detectable protein spots, and offered well‐resolved chain regions in comparison with those treated with DTT . Furthermore, this study introduced increasing the reduction power as a remedy to increase the reproducibility of two‐dimensional human tear proteome maps. In addition, the results of our assessment showed that improved reduction efficiency was accompanied by increased procedure reproducibility from 42% to 89%.