Isolation and solubilization of proteins after TRIzol® extraction of RNA and DNA from patient material following prolonged storage
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 transc...
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| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2007-04-01
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| Series: | BioTechniques |
| Online Access: | https://www.future-science.com/doi/10.2144/000112401 |
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| _version_ | 1850151974973997056 |
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| author | Amanda B. Hummon Sharlene R. Lim Michael J. Difilippantonio Thomas Ried |
| author_facet | Amanda B. Hummon Sharlene R. Lim Michael J. Difilippantonio Thomas Ried |
| author_sort | Amanda B. Hummon |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-55cb1b4f62eb45b885b9ebc503e01e50 |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2007-04-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-55cb1b4f62eb45b885b9ebc503e01e502025-08-20T02:26:06ZengTaylor & Francis GroupBioTechniques0736-62051940-98182007-04-0142446747210.2144/000112401Isolation and solubilization of proteins after TRIzol® extraction of RNA and DNA from patient material following prolonged storageAmanda B. Hummon0Sharlene R. Lim1Michael J. Difilippantonio2Thomas Ried31National Institutes of Health, Bethesda, MD2University of California, Berkeley, Berkeley, CA, USA1National Institutes of Health, Bethesda, MD1National Institutes of Health, Bethesda, MDA 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.https://www.future-science.com/doi/10.2144/000112401 |
| spellingShingle | Amanda B. Hummon Sharlene R. Lim Michael J. Difilippantonio Thomas Ried Isolation and solubilization of proteins after TRIzol® extraction of RNA and DNA from patient material following prolonged storage BioTechniques |
| title | Isolation and solubilization of proteins after TRIzol® extraction of RNA and DNA from patient material following prolonged storage |
| title_full | Isolation and solubilization of proteins after TRIzol® extraction of RNA and DNA from patient material following prolonged storage |
| title_fullStr | Isolation and solubilization of proteins after TRIzol® extraction of RNA and DNA from patient material following prolonged storage |
| title_full_unstemmed | Isolation and solubilization of proteins after TRIzol® extraction of RNA and DNA from patient material following prolonged storage |
| title_short | Isolation and solubilization of proteins after TRIzol® extraction of RNA and DNA from patient material following prolonged storage |
| title_sort | isolation and solubilization of proteins after trizol r extraction of rna and dna from patient material following prolonged storage |
| url | https://www.future-science.com/doi/10.2144/000112401 |
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