Comparison of the sensitivity of Western blotting between PVDF and NC membranes

Abstract Western blotting (WB) is one of the most widely used techniques to identify proteins as well as post translational modifications of proteins. The selection of electroblotted membrane is one of the key factors affecting the detection sensitivity of the protein which is transferred from gel t...

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Autores principales:	Yufang Xiang, Yuanyuan Zheng, Shaobo Liu, Gang Liu, Zhi Li, Weijie Dong
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/215473a333b3470ab9b167f88d266942
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spelling	oai:doaj.org-article:215473a333b3470ab9b167f88d2669422021-12-02T17:52:13ZComparison of the sensitivity of Western blotting between PVDF and NC membranes10.1038/s41598-021-91521-82045-2322https://doaj.org/article/215473a333b3470ab9b167f88d2669422021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-91521-8https://doaj.org/toc/2045-2322Abstract Western blotting (WB) is one of the most widely used techniques to identify proteins as well as post translational modifications of proteins. The selection of electroblotted membrane is one of the key factors affecting the detection sensitivity of the protein which is transferred from gel to membrane in WB. The most common used membranes are polyvinylidene fluoride (PVDF) and nitrocellulose (NC) membranes. Which membrane of these two is more suitable for WB has not been reported so far. Here, by incubating proteins which were transferred to PVDF or NC membranes with a series of antibodies and different types of lectins, we investigated the relationship between the binding ability of these two membranes to proteins or glycoproteins and the molecular weight of the target protein. The antibody re-probed ability of the two membranes was also explored. Moreover, we verified the above results by directly incubating proteins having different molecular weights onto PVDF or NC membranes. Bound proteins were stained with direct blue-71, and the staining intensity was quantitated by scanning and densitometry.Yufang XiangYuanyuan ZhengShaobo LiuGang LiuZhi LiWeijie DongNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yufang Xiang Yuanyuan Zheng Shaobo Liu Gang Liu Zhi Li Weijie Dong Comparison of the sensitivity of Western blotting between PVDF and NC membranes
description	Abstract Western blotting (WB) is one of the most widely used techniques to identify proteins as well as post translational modifications of proteins. The selection of electroblotted membrane is one of the key factors affecting the detection sensitivity of the protein which is transferred from gel to membrane in WB. The most common used membranes are polyvinylidene fluoride (PVDF) and nitrocellulose (NC) membranes. Which membrane of these two is more suitable for WB has not been reported so far. Here, by incubating proteins which were transferred to PVDF or NC membranes with a series of antibodies and different types of lectins, we investigated the relationship between the binding ability of these two membranes to proteins or glycoproteins and the molecular weight of the target protein. The antibody re-probed ability of the two membranes was also explored. Moreover, we verified the above results by directly incubating proteins having different molecular weights onto PVDF or NC membranes. Bound proteins were stained with direct blue-71, and the staining intensity was quantitated by scanning and densitometry.
format	article
author	Yufang Xiang Yuanyuan Zheng Shaobo Liu Gang Liu Zhi Li Weijie Dong
author_facet	Yufang Xiang Yuanyuan Zheng Shaobo Liu Gang Liu Zhi Li Weijie Dong
author_sort	Yufang Xiang
title	Comparison of the sensitivity of Western blotting between PVDF and NC membranes
title_short	Comparison of the sensitivity of Western blotting between PVDF and NC membranes
title_full	Comparison of the sensitivity of Western blotting between PVDF and NC membranes
title_fullStr	Comparison of the sensitivity of Western blotting between PVDF and NC membranes
title_full_unstemmed	Comparison of the sensitivity of Western blotting between PVDF and NC membranes
title_sort	comparison of the sensitivity of western blotting between pvdf and nc membranes
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/215473a333b3470ab9b167f88d266942
work_keys_str_mv	AT yufangxiang comparisonofthesensitivityofwesternblottingbetweenpvdfandncmembranes AT yuanyuanzheng comparisonofthesensitivityofwesternblottingbetweenpvdfandncmembranes AT shaoboliu comparisonofthesensitivityofwesternblottingbetweenpvdfandncmembranes AT gangliu comparisonofthesensitivityofwesternblottingbetweenpvdfandncmembranes AT zhili comparisonofthesensitivityofwesternblottingbetweenpvdfandncmembranes AT weijiedong comparisonofthesensitivityofwesternblottingbetweenpvdfandncmembranes
_version_	1718379214131953664

Comparison of the sensitivity of Western blotting between PVDF and NC membranes

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