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

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

Proteomics (2006-02-04)
Cristina-Maria Vâlcu, Katja Schlink
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Tributylphosphine, ≥93.5% (Tri-N-butylphosphine, GC)
Sigma-Aldrich
Tributylphosphine, mixture of isomers, 97%
Sigma-Aldrich
Tri-n-butylphosphine, 99%
Sigma-Aldrich
Tri-n-butylphosphine, 97%
Sigma-Aldrich
Tributylphosphine solution, 200 mM (in N-methyl-2-pyrrolidinone), liquid