Cargando…
Gel-Based and Gel-Free Quantitative Proteomics Approaches at a Glance
Two-dimensional gel electrophoresis (2-DE) is widely applied and remains the method of choice in proteomics; however, pervasive 2-DE-related concerns undermine its prospects as a dominant separation technique in proteome research. Consequently, the state-of-the-art shotgun techniques are slowly taki...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Hindawi Publishing Corporation
2012
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3508552/ https://www.ncbi.nlm.nih.gov/pubmed/23213324 http://dx.doi.org/10.1155/2012/494572 |
_version_ | 1782251203726409728 |
---|---|
author | Abdallah, Cosette Dumas-Gaudot, Eliane Renaut, Jenny Sergeant, Kjell |
author_facet | Abdallah, Cosette Dumas-Gaudot, Eliane Renaut, Jenny Sergeant, Kjell |
author_sort | Abdallah, Cosette |
collection | PubMed |
description | Two-dimensional gel electrophoresis (2-DE) is widely applied and remains the method of choice in proteomics; however, pervasive 2-DE-related concerns undermine its prospects as a dominant separation technique in proteome research. Consequently, the state-of-the-art shotgun techniques are slowly taking over and utilising the rapid expansion and advancement of mass spectrometry (MS) to provide a new toolbox of gel-free quantitative techniques. When coupled to MS, the shotgun proteomic pipeline can fuel new routes in sensitive and high-throughput profiling of proteins, leading to a high accuracy in quantification. Although label-based approaches, either chemical or metabolic, gained popularity in quantitative proteomics because of the multiplexing capacity, these approaches are not without drawbacks. The burgeoning label-free methods are tag independent and suitable for all kinds of samples. The challenges in quantitative proteomics are more prominent in plants due to difficulties in protein extraction, some protein abundance in green tissue, and the absence of well-annotated and completed genome sequences. The goal of this perspective assay is to present the balance between the strengths and weaknesses of the available gel-based and -free methods and their application to plants. The latest trends in peptide fractionation amenable to MS analysis are as well discussed. |
format | Online Article Text |
id | pubmed-3508552 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Hindawi Publishing Corporation |
record_format | MEDLINE/PubMed |
spelling | pubmed-35085522012-12-04 Gel-Based and Gel-Free Quantitative Proteomics Approaches at a Glance Abdallah, Cosette Dumas-Gaudot, Eliane Renaut, Jenny Sergeant, Kjell Int J Plant Genomics Review Article Two-dimensional gel electrophoresis (2-DE) is widely applied and remains the method of choice in proteomics; however, pervasive 2-DE-related concerns undermine its prospects as a dominant separation technique in proteome research. Consequently, the state-of-the-art shotgun techniques are slowly taking over and utilising the rapid expansion and advancement of mass spectrometry (MS) to provide a new toolbox of gel-free quantitative techniques. When coupled to MS, the shotgun proteomic pipeline can fuel new routes in sensitive and high-throughput profiling of proteins, leading to a high accuracy in quantification. Although label-based approaches, either chemical or metabolic, gained popularity in quantitative proteomics because of the multiplexing capacity, these approaches are not without drawbacks. The burgeoning label-free methods are tag independent and suitable for all kinds of samples. The challenges in quantitative proteomics are more prominent in plants due to difficulties in protein extraction, some protein abundance in green tissue, and the absence of well-annotated and completed genome sequences. The goal of this perspective assay is to present the balance between the strengths and weaknesses of the available gel-based and -free methods and their application to plants. The latest trends in peptide fractionation amenable to MS analysis are as well discussed. Hindawi Publishing Corporation 2012 2012-11-20 /pmc/articles/PMC3508552/ /pubmed/23213324 http://dx.doi.org/10.1155/2012/494572 Text en Copyright © 2012 Cosette Abdallah et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Review Article Abdallah, Cosette Dumas-Gaudot, Eliane Renaut, Jenny Sergeant, Kjell Gel-Based and Gel-Free Quantitative Proteomics Approaches at a Glance |
title | Gel-Based and Gel-Free Quantitative Proteomics Approaches at a Glance |
title_full | Gel-Based and Gel-Free Quantitative Proteomics Approaches at a Glance |
title_fullStr | Gel-Based and Gel-Free Quantitative Proteomics Approaches at a Glance |
title_full_unstemmed | Gel-Based and Gel-Free Quantitative Proteomics Approaches at a Glance |
title_short | Gel-Based and Gel-Free Quantitative Proteomics Approaches at a Glance |
title_sort | gel-based and gel-free quantitative proteomics approaches at a glance |
topic | Review Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3508552/ https://www.ncbi.nlm.nih.gov/pubmed/23213324 http://dx.doi.org/10.1155/2012/494572 |
work_keys_str_mv | AT abdallahcosette gelbasedandgelfreequantitativeproteomicsapproachesataglance AT dumasgaudoteliane gelbasedandgelfreequantitativeproteomicsapproachesataglance AT renautjenny gelbasedandgelfreequantitativeproteomicsapproachesataglance AT sergeantkjell gelbasedandgelfreequantitativeproteomicsapproachesataglance |