Protein extraction protocol from Musa sp. shoots and roots tissue for non-reducing one dimensional SDS-PAGE analysis

Background: Protein extraction from plant tissues is a great challenge since they contain low amount of proteins and rich in proteases, secondary metabolites and oxidative enzymes. Besides, dealing with limited amount of starting material would pose another challenge in getting high protein yield. T...

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Main Authors: Mohd Nasir, Nurul Najihah, Ho, Chai Ling, Lamasudin, Dhilia Udie, Saidi, Noor Baity
Format: Article
Language:English
Published: Bentham Science Publishers 2018
Online Access:http://psasir.upm.edu.my/id/eprint/73666/1/Protein%20extraction%20protocol%20from%20Musa%20sp.%20shoots%20and%20roots%20tissue%20for%20non-reducing%20one%20dimensional%20SDS-PAGE%20analysis.pdf
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author Mohd Nasir, Nurul Najihah
Ho, Chai Ling
Lamasudin, Dhilia Udie
Saidi, Noor Baity
author_facet Mohd Nasir, Nurul Najihah
Ho, Chai Ling
Lamasudin, Dhilia Udie
Saidi, Noor Baity
author_sort Mohd Nasir, Nurul Najihah
collection UPM
description Background: Protein extraction from plant tissues is a great challenge since they contain low amount of proteins and rich in proteases, secondary metabolites and oxidative enzymes. Besides, dealing with limited amount of starting material would pose another challenge in getting high protein yield. The aim of this study is to determine the best protein extraction protocol for recalcitrant tissues of banana (Musa sp.) specific for non-reducing one dimensional SDS-PAGE analysis. Methods: Three protein extraction protocols were compared which include TCA-acetone, phenol and phosphate buffer saline (PBS) using small amount of starting material. Modifications were done to each protocol to suit non-reducing SDS-PAGE analysis and subsequent downstream processes. Results: Of the three protocols compared, TCA-acetone protocol gave the highest yield and quality of protein extract from only a small amount of starting material based on protein quantification and non-reducing one dimensional SDS-PAGE analysis compared to phenol and phosphate buffer saline (PBS) protocol. Conclusion: Even though various protein extraction protocols have been developed to date, there is no one-size-fits-all when it comes to different types of plant tissues from varieties of plant species. TCA-acetone protocol was found to be most effective in extracting protein from banana shoot and root tissues even for small amount of starting material in absence of reducing agent.
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spelling upm.eprints-736662020-06-24T03:46:02Z http://psasir.upm.edu.my/id/eprint/73666/ Protein extraction protocol from Musa sp. shoots and roots tissue for non-reducing one dimensional SDS-PAGE analysis Mohd Nasir, Nurul Najihah Ho, Chai Ling Lamasudin, Dhilia Udie Saidi, Noor Baity Background: Protein extraction from plant tissues is a great challenge since they contain low amount of proteins and rich in proteases, secondary metabolites and oxidative enzymes. Besides, dealing with limited amount of starting material would pose another challenge in getting high protein yield. The aim of this study is to determine the best protein extraction protocol for recalcitrant tissues of banana (Musa sp.) specific for non-reducing one dimensional SDS-PAGE analysis. Methods: Three protein extraction protocols were compared which include TCA-acetone, phenol and phosphate buffer saline (PBS) using small amount of starting material. Modifications were done to each protocol to suit non-reducing SDS-PAGE analysis and subsequent downstream processes. Results: Of the three protocols compared, TCA-acetone protocol gave the highest yield and quality of protein extract from only a small amount of starting material based on protein quantification and non-reducing one dimensional SDS-PAGE analysis compared to phenol and phosphate buffer saline (PBS) protocol. Conclusion: Even though various protein extraction protocols have been developed to date, there is no one-size-fits-all when it comes to different types of plant tissues from varieties of plant species. TCA-acetone protocol was found to be most effective in extracting protein from banana shoot and root tissues even for small amount of starting material in absence of reducing agent. Bentham Science Publishers 2018 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/73666/1/Protein%20extraction%20protocol%20from%20Musa%20sp.%20shoots%20and%20roots%20tissue%20for%20non-reducing%20one%20dimensional%20SDS-PAGE%20analysis.pdf Mohd Nasir, Nurul Najihah and Ho, Chai Ling and Lamasudin, Dhilia Udie and Saidi, Noor Baity (2018) Protein extraction protocol from Musa sp. shoots and roots tissue for non-reducing one dimensional SDS-PAGE analysis. Natural Products Journal, 8 (3). 191 - 195. ISSN 2210-3155; ESSN: 2210-3163 http://www.eurekaselect.com/node/154392/article/protein-extraction-protocol-from-musa-sp-shoots-and-roots-tissue-for-non-reducing-one-dimensional-sds-page-analysis 10.2174/2210315507666170724111327
spellingShingle Mohd Nasir, Nurul Najihah
Ho, Chai Ling
Lamasudin, Dhilia Udie
Saidi, Noor Baity
Protein extraction protocol from Musa sp. shoots and roots tissue for non-reducing one dimensional SDS-PAGE analysis
title Protein extraction protocol from Musa sp. shoots and roots tissue for non-reducing one dimensional SDS-PAGE analysis
title_full Protein extraction protocol from Musa sp. shoots and roots tissue for non-reducing one dimensional SDS-PAGE analysis
title_fullStr Protein extraction protocol from Musa sp. shoots and roots tissue for non-reducing one dimensional SDS-PAGE analysis
title_full_unstemmed Protein extraction protocol from Musa sp. shoots and roots tissue for non-reducing one dimensional SDS-PAGE analysis
title_short Protein extraction protocol from Musa sp. shoots and roots tissue for non-reducing one dimensional SDS-PAGE analysis
title_sort protein extraction protocol from musa sp shoots and roots tissue for non reducing one dimensional sds page analysis
url http://psasir.upm.edu.my/id/eprint/73666/1/Protein%20extraction%20protocol%20from%20Musa%20sp.%20shoots%20and%20roots%20tissue%20for%20non-reducing%20one%20dimensional%20SDS-PAGE%20analysis.pdf
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