2012
DOI: 10.1590/s1415-47572012000200016
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Abstract: Sample preparation is a critical step in two-dimensional gel electrophoresis (2-DE) of plant tissues. Here we describe a phenol/SDS procedure that, although greatly simplified, produced well-resolved and reproducible 2-DE profiles of protein extracts from soybean [Glycine max (L.) Merril] roots. Extractions were made in three replicates using both the original and simplified procedure. To evaluate the quality of the extracted proteins, ten spots were randomly selected and identified by mass spectrometry (MS). … Show more

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Cited by 28 publications
(15 citation statements)
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References 15 publications
(26 reference statements)
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“…Whole-cell proteins of soybean roots were extracted, from both the inoculated and non-inoculated treatments, following the simplified method described by Rodrigues et al [71]. IPG strips (pH 4–7, 13 cm, GE Healthcare, Uppsala, Sweden) were rehydrated overnight with aliquots of 350 μg of solubilized proteins.…”
Section: Methodsmentioning
confidence: 99%
“…Although there are several sample preparation protocols available in the literature (Palma et al 2011, Rodrigues et al 2012, protocols specifically designed for tomato species (Solanum lycopersicum L.) are not well defined.…”
Section: Introductionmentioning
confidence: 99%
“…Because of the biochemical heterogeneity, large amounts of proteases and oxidative enzymes, and the presence of non-protein contaminants can disturb the subsequent steps in proteomic approaches (Boonmee et al, 2011;Witzel et al, 2011). Therefore, there is not a universal protocol for extraction or a solvent that can capture an entire proteome, and adjustments to previously described protocols are required (Prinsi et al, 2011;Chatterjee et al, 2012;Rodrigues et al, 2012a;Zhang et al, 2012).…”
Section: Introductionmentioning
confidence: 99%
“…Although a single-step process for protein extraction would be highly desirable, no unique sample preparation method can be used to 2-DE analysis (Dunn, 1999). Therefore, many researchers developed and optimized some efficient methods such as a phenol/SDS-based method (Wang et al, 2006; Rodrigues et al, 2012) and non-phenol-based methods (Guerreiro et al, 1997; Natera et al, 2000), to find a simple method that could be applied regularly to proteomics studies of symbiosis interactions.…”
Section: Introductionmentioning
confidence: 99%
“…Furthermore, while interfering compounds are challenging in all plant tissues, they are particularly abundant in green (i.e., photosynthetically active) tissues . As a result, numerous protein extraction methods using various combinations of acetone, methanol, chloroform, phenol, detergents, and molecular weight cutoff filters have been evaluated on plant tissues . However, nearly all the evaluations of extraction methods have examined compatibility with 2‐DE rather than reverse phase LC, which is necessary for deep and quantitative proteome profiling.…”
mentioning
confidence: 99%