An efficient solubilization buffer for plant proteins focused in immobilized pH gradients.

Abstract:

:The solubilization of a large array of proteins before electrophoresis itself is a very critical point for proteomic analyses. We compared the efficiency of several different solubilization buffers. From this work, we defined a very efficient solubilization buffer, including two chaotropes, two reducing agents (R2), two detergents (D2), and two kinds of carrier ampholytes in combination. This so-called R2D2 buffer (5 M urea, 2 M thiourea, 2% 3-[(3-cholamidopropyl) dimethyl-ammonio]-1-propane-sulfonate, 2% N-decyl-N,N-dimethyl-3-ammonio-1-propane-sulfonate, 20 mM dithiothreitol, 5 mM Tris(2-carboxyethyl) phosphine, 0.5% carrier ampholytes 4-6.5, 0.25% carrier ampholytes 3-10) proved to be very efficient for a large range of different samples and allowed us to obtain two-dimensional gels of high resolution and quality.

journal_name

Proteomics

journal_title

Proteomics

authors

Méchin V,Consoli L,Le Guilloux M,Damerval C

doi

10.1002/pmic.200300450

subject

Has Abstract

pub_date

2003-07-01 00:00:00

pages

1299-302

issue

7

eissn

1615-9853

issn

1615-9861

journal_volume

3

pub_type

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