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Reduction of proteins during sample preparation and two‐dimensional gel electrophoresis of woody plant samples
Author(s) -
Vâlcu CristinaMaria,
Schlink Katja
Publication year - 2006
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200500314
Subject(s) - tcep , chemistry , chromatography , phosphine , extraction (chemistry) , tris , reducing agent , sample preparation , protein purification , beech , biochemistry , botany , organic chemistry , biology , catalysis
Protein extraction procedure and the reducing agent content (DTT, dithioerythritol, tributyl phosphine and tris (2‐carboxyethyl) phosphine (TCEP)) of the sample and rehydration buffers were optimised for European beech leaves and roots and Norway spruce needles. Optimal extraction was achieved with 100 mM DTT for leaves and needles and a mixture of 2 mM TCEP and 50 mM DTT for roots. Performing IEF in buffers containing hydroxyethyldisulphide significantly enhanced the quality of separation for all proteins except for acidic root proteins, which were optimally focused in the same buffer as extracted.