Tissue-Specific Microdissection Coupled with ProteinChip® Array Technologies: Applications in Cancer Research
Analysis of whole genomes to monitor specific changes in gene activation or changes in gene copy number due to perturbation has recently become possible using DNA chip technologies. It is now becoming apparent, however, that knowing the genetic sequence encoding a protein is not sufficient to predic...
Saved in:
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Taylor & Francis Group
2000-11-01
|
| Series: | BioTechniques |
| Online Access: | https://www.future-science.com/doi/10.2144/00295rr02 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850151974420348928 |
|---|---|
| author | F. von Eggeling H. Davies L. Lomas W. Fiedler K. Junker U. Claussen G. Ernst |
| author_facet | F. von Eggeling H. Davies L. Lomas W. Fiedler K. Junker U. Claussen G. Ernst |
| author_sort | F. von Eggeling |
| collection | DOAJ |
| description | Analysis of whole genomes to monitor specific changes in gene activation or changes in gene copy number due to perturbation has recently become possible using DNA chip technologies. It is now becoming apparent, however, that knowing the genetic sequence encoding a protein is not sufficient to predict the size or biological nature of a protein. This can be particularly important in cancer research where posttranslational modifications of a protein can specifically lead to the disease. To address this area, several proteomic tools have been developed. Currently the most widely used proteomics tool is two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), which can display protein expression patterns to a high degree of resolution. However, 2D-PAGE can be time consuming; the analysis is complicated and, compared with DNA techniques, is not very sensitive. Although some of these problems can be alleviated by using high-quality homogeneous samples, such as those generated using microdissection techniques, the quantity of sample is often limited and may take several days to generate sufficient material for a single 2D-PAGE analysis. As an alternative to 2D-PAGE, a preliminary study using a new technique was used to generate protein expression patterns from either whole tissue extracts or microdissected material. Surface-enhanced laser desorption and ionization allows the retention of proteins on a solid-phase chromatographic surface or ProteinChip® Array with direct detection of retained proteins by time-of-flight mass spectrometry. Using this system, we analyzed tumor and normal tissue from head and neck cancer and microdissected melanoma to determine differentially expressed proteins. In particular, comparisons of the protein expression patterns from microdissected normal and tumor tissues indicated several differences, highlighting the importance of extremely defined tissue lysates for protein profiling. |
| format | Article |
| id | doaj-art-547aef6478d54ebe8b8cb29e7b71eeb1 |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2000-11-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-547aef6478d54ebe8b8cb29e7b71eeb12025-08-20T02:26:06ZengTaylor & Francis GroupBioTechniques0736-62051940-98182000-11-012951066107010.2144/00295rr02Tissue-Specific Microdissection Coupled with ProteinChip® Array Technologies: Applications in Cancer ResearchF. von Eggeling0H. Davies1L. Lomas2W. Fiedler3K. Junker4U. Claussen5G. Ernst61Institute of Human Genetics and Anthropology, Jena, UK1Institute of Human Genetics and Anthropology, Jena, UK1Institute of Human Genetics and Anthropology, Jena, UK1Institute of Human Genetics and Anthropology, Jena, UK1Institute of Human Genetics and Anthropology, Jena, UK1Institute of Human Genetics and Anthropology, Jena, UK1Institute of Human Genetics and Anthropology, Jena, UKAnalysis of whole genomes to monitor specific changes in gene activation or changes in gene copy number due to perturbation has recently become possible using DNA chip technologies. It is now becoming apparent, however, that knowing the genetic sequence encoding a protein is not sufficient to predict the size or biological nature of a protein. This can be particularly important in cancer research where posttranslational modifications of a protein can specifically lead to the disease. To address this area, several proteomic tools have been developed. Currently the most widely used proteomics tool is two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), which can display protein expression patterns to a high degree of resolution. However, 2D-PAGE can be time consuming; the analysis is complicated and, compared with DNA techniques, is not very sensitive. Although some of these problems can be alleviated by using high-quality homogeneous samples, such as those generated using microdissection techniques, the quantity of sample is often limited and may take several days to generate sufficient material for a single 2D-PAGE analysis. As an alternative to 2D-PAGE, a preliminary study using a new technique was used to generate protein expression patterns from either whole tissue extracts or microdissected material. Surface-enhanced laser desorption and ionization allows the retention of proteins on a solid-phase chromatographic surface or ProteinChip® Array with direct detection of retained proteins by time-of-flight mass spectrometry. Using this system, we analyzed tumor and normal tissue from head and neck cancer and microdissected melanoma to determine differentially expressed proteins. In particular, comparisons of the protein expression patterns from microdissected normal and tumor tissues indicated several differences, highlighting the importance of extremely defined tissue lysates for protein profiling.https://www.future-science.com/doi/10.2144/00295rr02 |
| spellingShingle | F. von Eggeling H. Davies L. Lomas W. Fiedler K. Junker U. Claussen G. Ernst Tissue-Specific Microdissection Coupled with ProteinChip® Array Technologies: Applications in Cancer Research BioTechniques |
| title | Tissue-Specific Microdissection Coupled with ProteinChip® Array Technologies: Applications in Cancer Research |
| title_full | Tissue-Specific Microdissection Coupled with ProteinChip® Array Technologies: Applications in Cancer Research |
| title_fullStr | Tissue-Specific Microdissection Coupled with ProteinChip® Array Technologies: Applications in Cancer Research |
| title_full_unstemmed | Tissue-Specific Microdissection Coupled with ProteinChip® Array Technologies: Applications in Cancer Research |
| title_short | Tissue-Specific Microdissection Coupled with ProteinChip® Array Technologies: Applications in Cancer Research |
| title_sort | tissue specific microdissection coupled with proteinchip r array technologies applications in cancer research |
| url | https://www.future-science.com/doi/10.2144/00295rr02 |
| work_keys_str_mv | AT fvoneggeling tissuespecificmicrodissectioncoupledwithproteinchiparraytechnologiesapplicationsincancerresearch AT hdavies tissuespecificmicrodissectioncoupledwithproteinchiparraytechnologiesapplicationsincancerresearch AT llomas tissuespecificmicrodissectioncoupledwithproteinchiparraytechnologiesapplicationsincancerresearch AT wfiedler tissuespecificmicrodissectioncoupledwithproteinchiparraytechnologiesapplicationsincancerresearch AT kjunker tissuespecificmicrodissectioncoupledwithproteinchiparraytechnologiesapplicationsincancerresearch AT uclaussen tissuespecificmicrodissectioncoupledwithproteinchiparraytechnologiesapplicationsincancerresearch AT gernst tissuespecificmicrodissectioncoupledwithproteinchiparraytechnologiesapplicationsincancerresearch |