Determination of Green Fluorescent Protein by Ultraviolet-Excited Gel Imaging

The green fluorescent protein has gained interest in bioanalytical applications due to its visible fluorescence. As the usage of green fluorescent protein increases, more appropriate fluorescence instrumentation is required. Most fluorescence instrumentation uses ultraviolet light as the excitation...

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Main Authors: Hor, Chee Heng, Chew, Few Ne
Format: Article
Language:English
Published: Taylor & Francis 2016
Subjects:
Online Access:http://umpir.ump.edu.my/id/eprint/14577/7/fkksa-2016-chew-Determination%20Of%20Green%20Fluorescent%20Protein.pdf
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author Hor, Chee Heng
Chew, Few Ne
author_facet Hor, Chee Heng
Chew, Few Ne
author_sort Hor, Chee Heng
collection UMP
description The green fluorescent protein has gained interest in bioanalytical applications due to its visible fluorescence. As the usage of green fluorescent protein increases, more appropriate fluorescence instrumentation is required. Most fluorescence instrumentation uses ultraviolet light as the excitation source for the determination of green fluorescent protein. However, ultraviolet radiation may damage biological molecules and affect the quantitative analysis. In this study, the effects of the ultraviolet radiation period and the mass of green fluorescent protein on the fluorescence determination were characterized using gel imaging. The ultraviolet illumination period affected the green fluorescent protein fluorescence intensity. The fluorescence increased with the ultraviolet illumination time from 30–90 s. However, the fluorescence intensity decreased when the excitation period was longer, probably due to photobleaching. The photobleaching decreased when a higher concentration of enhanced green fluorescent protein was employed. This gel imaging study has provided a better understanding of the optimum conditions for the determination of green fluorescent protein.
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spelling UMPir145772017-12-15T06:44:55Z http://umpir.ump.edu.my/id/eprint/14577/ Determination of Green Fluorescent Protein by Ultraviolet-Excited Gel Imaging Hor, Chee Heng Chew, Few Ne TP Chemical technology The green fluorescent protein has gained interest in bioanalytical applications due to its visible fluorescence. As the usage of green fluorescent protein increases, more appropriate fluorescence instrumentation is required. Most fluorescence instrumentation uses ultraviolet light as the excitation source for the determination of green fluorescent protein. However, ultraviolet radiation may damage biological molecules and affect the quantitative analysis. In this study, the effects of the ultraviolet radiation period and the mass of green fluorescent protein on the fluorescence determination were characterized using gel imaging. The ultraviolet illumination period affected the green fluorescent protein fluorescence intensity. The fluorescence increased with the ultraviolet illumination time from 30–90 s. However, the fluorescence intensity decreased when the excitation period was longer, probably due to photobleaching. The photobleaching decreased when a higher concentration of enhanced green fluorescent protein was employed. This gel imaging study has provided a better understanding of the optimum conditions for the determination of green fluorescent protein. Taylor & Francis 2016-04-11 Article PeerReviewed application/pdf en http://umpir.ump.edu.my/id/eprint/14577/7/fkksa-2016-chew-Determination%20Of%20Green%20Fluorescent%20Protein.pdf Hor, Chee Heng and Chew, Few Ne (2016) Determination of Green Fluorescent Protein by Ultraviolet-Excited Gel Imaging. Instrumentation Science and Technology, 44 (5). pp. 558-565. ISSN 1525-6030. (Published) http://dx.doi.org/10.1080/10739149.2016.1170032 DOI: 10.1080/10739149.2016.1170032
spellingShingle TP Chemical technology
Hor, Chee Heng
Chew, Few Ne
Determination of Green Fluorescent Protein by Ultraviolet-Excited Gel Imaging
title Determination of Green Fluorescent Protein by Ultraviolet-Excited Gel Imaging
title_full Determination of Green Fluorescent Protein by Ultraviolet-Excited Gel Imaging
title_fullStr Determination of Green Fluorescent Protein by Ultraviolet-Excited Gel Imaging
title_full_unstemmed Determination of Green Fluorescent Protein by Ultraviolet-Excited Gel Imaging
title_short Determination of Green Fluorescent Protein by Ultraviolet-Excited Gel Imaging
title_sort determination of green fluorescent protein by ultraviolet excited gel imaging
topic TP Chemical technology
url http://umpir.ump.edu.my/id/eprint/14577/7/fkksa-2016-chew-Determination%20Of%20Green%20Fluorescent%20Protein.pdf
work_keys_str_mv AT horcheeheng determinationofgreenfluorescentproteinbyultravioletexcitedgelimaging
AT chewfewne determinationofgreenfluorescentproteinbyultravioletexcitedgelimaging