Optimization of 2-dimensional gel electrophoresis for proteomic studies of oil palm roots
2D electrophoresis (2-DE) is a proteomic tool that has been widely used as this technique is capable of resolving and separating thousands of proteins in a single gel for parallel analysis. 2-DE allows the identification of individual protein and understanding of the host...
| Main Author: | |
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| Format: | Project Paper Report |
| Language: | English |
| Published: |
2015
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| Online Access: | http://psasir.upm.edu.my/id/eprint/91044/1/FBSB%202015%20154%20-%20IR.pdf |
| _version_ | 1825936947690864640 |
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| author | Nik Zuriadi, Nik Nur A'in |
| author_facet | Nik Zuriadi, Nik Nur A'in |
| author_sort | Nik Zuriadi, Nik Nur A'in |
| collection | UPM |
| description | 2D electrophoresis (2-DE) is a proteomic tool that has been widely used as this
technique is capable of resolving and separating thousands of proteins in a single gel for
parallel analysis. 2-DE allows the identification of individual protein and understanding
of the host-pathogen interaction during their molecular crosstalks. In addition, this
modification technique also allows the study of protein profiling of solid biological
samples. However, the analytical conditions need to be optimized to achieve optimal parameters and
procedures for 2-D gel images. In the present study, optimization was carried out which aimed to
improve image quality, reproducibility and sensitivity of 2-D gel. Generally, focusing time
and voltage of Isoelectric focusing (IEF) affect the reproducibility of 2-D gel images
for protein analysis. It was found that increasing voltage and time for IEF running were optimal
for 2-DE. Reducing development time, enhancing the washing step, prolong fixation to
overnight and the use of freshly prepared reagents of silver staining produced better
image resolution and enhanced sensitivity. |
| first_indexed | 2024-03-06T10:52:03Z |
| format | Project Paper Report |
| id | upm.eprints-91044 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T10:52:03Z |
| publishDate | 2015 |
| record_format | dspace |
| spelling | upm.eprints-910442021-10-26T00:52:27Z http://psasir.upm.edu.my/id/eprint/91044/ Optimization of 2-dimensional gel electrophoresis for proteomic studies of oil palm roots Nik Zuriadi, Nik Nur A'in 2D electrophoresis (2-DE) is a proteomic tool that has been widely used as this technique is capable of resolving and separating thousands of proteins in a single gel for parallel analysis. 2-DE allows the identification of individual protein and understanding of the host-pathogen interaction during their molecular crosstalks. In addition, this modification technique also allows the study of protein profiling of solid biological samples. However, the analytical conditions need to be optimized to achieve optimal parameters and procedures for 2-D gel images. In the present study, optimization was carried out which aimed to improve image quality, reproducibility and sensitivity of 2-D gel. Generally, focusing time and voltage of Isoelectric focusing (IEF) affect the reproducibility of 2-D gel images for protein analysis. It was found that increasing voltage and time for IEF running were optimal for 2-DE. Reducing development time, enhancing the washing step, prolong fixation to overnight and the use of freshly prepared reagents of silver staining produced better image resolution and enhanced sensitivity. 2015-06 Project Paper Report NonPeerReviewed text en http://psasir.upm.edu.my/id/eprint/91044/1/FBSB%202015%20154%20-%20IR.pdf Nik Zuriadi, Nik Nur A'in (2015) Optimization of 2-dimensional gel electrophoresis for proteomic studies of oil palm roots. [Project Paper Report] |
| spellingShingle | Nik Zuriadi, Nik Nur A'in Optimization of 2-dimensional gel electrophoresis for proteomic studies of oil palm roots |
| title | Optimization of 2-dimensional gel electrophoresis for proteomic studies of oil palm roots |
| title_full | Optimization of 2-dimensional gel electrophoresis for proteomic studies of oil palm roots |
| title_fullStr | Optimization of 2-dimensional gel electrophoresis for proteomic studies of oil palm roots |
| title_full_unstemmed | Optimization of 2-dimensional gel electrophoresis for proteomic studies of oil palm roots |
| title_short | Optimization of 2-dimensional gel electrophoresis for proteomic studies of oil palm roots |
| title_sort | optimization of 2 dimensional gel electrophoresis for proteomic studies of oil palm roots |
| url | http://psasir.upm.edu.my/id/eprint/91044/1/FBSB%202015%20154%20-%20IR.pdf |
| work_keys_str_mv | AT nikzuriadiniknurain optimizationof2dimensionalgelelectrophoresisforproteomicstudiesofoilpalmroots |