Isolation and solubilization of proteins after TRIzol® extraction of RNA and DNA from patient material following prolonged storage
| Autor: | Amanda B. Hummon, Sharlene R. Lim, Michael J. Difilippantonio, Thomas Ried |
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| Jazyk: | angličtina |
| Rok vydání: | 2007 |
| Předmět: | |
| Zdroj: | BioTechniques, Vol 42, Iss 4, Pp 467-472 (2007) |
| Druh dokumentu: | article |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/000112401 |
| Popis: | A systems approach is being applied in many areas of the biological sciences, particularly in cancer research. The coordinated, simultaneous extraction of DNA, RNA, and proteins from a single sample is crucial for accurate correlations between genomic aberrations and their consequences on the transcriptome and proteome. We present an approach to extract and completely solubilize up to 98% of the total protein recovered from archived samples following TRIzol® isolation of RNA and DNA. We also demonstrate using polyacrylamide gel electrophoresis (PAGE) and Western blot analysis that the proteins, representing both a wide molecular weight range and some posttranslational modifications, such as protein phosphorylation, remain stable in phenol-ethanol for up to 3 years at −20°C. |
| Databáze: | Directory of Open Access Journals |
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