A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots
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. Ext...
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| Format: | Article |
| Language: | English |
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Sociedade Brasileira de Genética
2012-01-01
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| Series: | Genetics and Molecular Biology |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572012000200016 |
| _version_ | 1818959284844625920 |
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| author | Elisete Pains Rodrigues Adalgisa Ribeiro Torres Jesiane Stefânia da Silva Batista Luciano Huergo Mariangela Hungria |
| author_facet | Elisete Pains Rodrigues Adalgisa Ribeiro Torres Jesiane Stefânia da Silva Batista Luciano Huergo Mariangela Hungria |
| author_sort | Elisete Pains Rodrigues |
| collection | DOAJ |
| description | 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). The 2-DE gels were equally well resolved, with no streaks or smears, and no significant differences were observed in protein yield, reproducibility, resolution or number of spots. Mass spectra of the ten selected spots were compared with database entries and allowed high-quality identification of proteins. The simplified protocol described here presents considerable savings of time and reagents without compromising the quality of 2-DE protein profiles and compatibility with MS analysis, and may facilitate the progress of proteomics studies of legume-rhizobia interactions. |
| first_indexed | 2024-12-20T11:39:12Z |
| format | Article |
| id | doaj.art-3b5a5a465acf4423850e53522bb2a180 |
| institution | Directory Open Access Journal |
| issn | 1415-4757 1678-4685 |
| language | English |
| last_indexed | 2024-12-20T11:39:12Z |
| publishDate | 2012-01-01 |
| publisher | Sociedade Brasileira de Genética |
| record_format | Article |
| series | Genetics and Molecular Biology |
| spelling | doaj.art-3b5a5a465acf4423850e53522bb2a1802022-12-21T19:42:02ZengSociedade Brasileira de GenéticaGenetics and Molecular Biology1415-47571678-46852012-01-0135134835210.1590/S1415-47572012000200016A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean rootsElisete Pains RodriguesAdalgisa Ribeiro TorresJesiane Stefânia da Silva BatistaLuciano HuergoMariangela HungriaSample 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). The 2-DE gels were equally well resolved, with no streaks or smears, and no significant differences were observed in protein yield, reproducibility, resolution or number of spots. Mass spectra of the ten selected spots were compared with database entries and allowed high-quality identification of proteins. The simplified protocol described here presents considerable savings of time and reagents without compromising the quality of 2-DE protein profiles and compatibility with MS analysis, and may facilitate the progress of proteomics studies of legume-rhizobia interactions.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-475720120002000162-DEprotein extractionproteomicssoybean rootssymbiosis |
| spellingShingle | Elisete Pains Rodrigues Adalgisa Ribeiro Torres Jesiane Stefânia da Silva Batista Luciano Huergo Mariangela Hungria A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots Genetics and Molecular Biology 2-DE protein extraction proteomics soybean roots symbiosis |
| title | A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots |
| title_full | A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots |
| title_fullStr | A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots |
| title_full_unstemmed | A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots |
| title_short | A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots |
| title_sort | simple economical and reproducible protein extraction protocol for proteomics studies of soybean roots |
| topic | 2-DE protein extraction proteomics soybean roots symbiosis |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572012000200016 |
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