Abstract
In any proteomic studies, protein extraction and sample preparation are the most crucial steps for obtaining optimal results. This is to ensure extracted proteins are not only high in yield but also clean from contaminants that could affect downstream proteomic applications such as two dimensional gel electrophoresis (2-DE) and mass spectrometry. Tissues from plants and trees such as Swietenia macrophylla are often rich in non-protein contaminating substances, which could interfere in the proteomic applications. S. macrophylla or also known as the mahogany is one of the most valuable tree species in the world. Studies on proteins for this tree as well as its seeds are very limited. We have extracted proteins from S. macrophylla seeds (specifically embryo tissues) using three different methods, each having different lysis buffer recipes. Furthermore, another set of samples were precipitated using trichloroacetic acid/acetone prior to the three extraction methods to further purify the protein samples. The results from 2-DE analysis showed approximately 240 protein spots were detected from the successful protocol using a lysis buffer of 9 M urea, 4% CHAPS, 0.5% triton X-100 and 100 mM DTT without TCA/acetone precipitation. This study highlights the aspects of sample preparation for S. macrophylla embryos, focusing on the total protein extraction and resolution in SDS-PAGE as well as 2-DE. Furthermore, this is the very first report of the proteome 2DE profile from S. macrophylla embryo.
| Original language | English |
|---|---|
| Pages (from-to) | 176-182 |
| Number of pages | 7 |
| Journal | Plant OMICS |
| Volume | 10 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - 1 Jul 2017 |
Fingerprint
Keywords
- Embryo
- Mahogany
- Proteomic
- Seeds
- Two-dimensional gel electrophoresis
ASJC Scopus subject areas
- Agronomy and Crop Science
- Plant Science
Cite this
A simple protein extraction method for proteomic analysis of mahogany (Swietenia macrophylla) embryos. / Alias, Noraliza; Wan Kamaruddin, Wan Mohd Aizat; Amin, Nor Datiakma Mat; Muhammad, Norwati; Mohd. Noor, Normah.
In: Plant OMICS, Vol. 10, No. 4, 01.07.2017, p. 176-182.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - A simple protein extraction method for proteomic analysis of mahogany (Swietenia macrophylla) embryos
AU - Alias, Noraliza
AU - Wan Kamaruddin, Wan Mohd Aizat
AU - Amin, Nor Datiakma Mat
AU - Muhammad, Norwati
AU - Mohd. Noor, Normah
PY - 2017/7/1
Y1 - 2017/7/1
N2 - In any proteomic studies, protein extraction and sample preparation are the most crucial steps for obtaining optimal results. This is to ensure extracted proteins are not only high in yield but also clean from contaminants that could affect downstream proteomic applications such as two dimensional gel electrophoresis (2-DE) and mass spectrometry. Tissues from plants and trees such as Swietenia macrophylla are often rich in non-protein contaminating substances, which could interfere in the proteomic applications. S. macrophylla or also known as the mahogany is one of the most valuable tree species in the world. Studies on proteins for this tree as well as its seeds are very limited. We have extracted proteins from S. macrophylla seeds (specifically embryo tissues) using three different methods, each having different lysis buffer recipes. Furthermore, another set of samples were precipitated using trichloroacetic acid/acetone prior to the three extraction methods to further purify the protein samples. The results from 2-DE analysis showed approximately 240 protein spots were detected from the successful protocol using a lysis buffer of 9 M urea, 4% CHAPS, 0.5% triton X-100 and 100 mM DTT without TCA/acetone precipitation. This study highlights the aspects of sample preparation for S. macrophylla embryos, focusing on the total protein extraction and resolution in SDS-PAGE as well as 2-DE. Furthermore, this is the very first report of the proteome 2DE profile from S. macrophylla embryo.
AB - In any proteomic studies, protein extraction and sample preparation are the most crucial steps for obtaining optimal results. This is to ensure extracted proteins are not only high in yield but also clean from contaminants that could affect downstream proteomic applications such as two dimensional gel electrophoresis (2-DE) and mass spectrometry. Tissues from plants and trees such as Swietenia macrophylla are often rich in non-protein contaminating substances, which could interfere in the proteomic applications. S. macrophylla or also known as the mahogany is one of the most valuable tree species in the world. Studies on proteins for this tree as well as its seeds are very limited. We have extracted proteins from S. macrophylla seeds (specifically embryo tissues) using three different methods, each having different lysis buffer recipes. Furthermore, another set of samples were precipitated using trichloroacetic acid/acetone prior to the three extraction methods to further purify the protein samples. The results from 2-DE analysis showed approximately 240 protein spots were detected from the successful protocol using a lysis buffer of 9 M urea, 4% CHAPS, 0.5% triton X-100 and 100 mM DTT without TCA/acetone precipitation. This study highlights the aspects of sample preparation for S. macrophylla embryos, focusing on the total protein extraction and resolution in SDS-PAGE as well as 2-DE. Furthermore, this is the very first report of the proteome 2DE profile from S. macrophylla embryo.
KW - Embryo
KW - Mahogany
KW - Proteomic
KW - Seeds
KW - Two-dimensional gel electrophoresis
UR - http://www.scopus.com/inward/record.url?scp=85028725397&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85028725397&partnerID=8YFLogxK
U2 - 10.21475/poj.10.04.17.pne323
DO - 10.21475/poj.10.04.17.pne323
M3 - Article
VL - 10
SP - 176
EP - 182
JO - Plant OMICS
JF - Plant OMICS
SN - 1836-0661
IS - 4
ER -