Reduction of proteins during sample preparation and two-dimensional gel electrophoresis of woody plant samples

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...

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Bibliographic Details
Published in:Proteomics (Weinheim) 2006-03, Vol.6 (5), p.1599-1605
Main Authors: Vâlcu, Cristina-Maria, Schlink, Katja
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Buffers
Dithiothreitol - chemistry
Electrophoresis, Gel, Two-Dimensional - methods
Fagus - chemistry
Fundamental and applied biological sciences. Psychology
Indicators and Reagents - chemistry
Miscellaneous
Oxidation-Reduction
Phosphines - chemistry
Picea - chemistry
Picea abies
Plant Proteins - chemistry
Plant Structures - chemistry
Proteins
Reducing agent
Sample preparation
Two-dimensional gel electrophoresis
Woody plant
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Summary: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.
ISSN:1615-9853
1615-9861
DOI:10.1002/pmic.200500314