Evaluation of UBE3A antibodies in mice and human cerebral organoids

Abstract UBE3A is an E3 ubiquitin ligase encoded by the neurally imprinted UBE3A gene. The abundance and subcellular distribution of UBE3A has been the topic of many previous studies as its dosage and localization has been linked to neurodevelopmental disorders including Autism, Dup15q syndrome, and...

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Autores principales:	Dilara Sen, Zuzana Drobna, Albert J. Keung
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9e387ac290914899a72d3d48062a092f
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spelling	oai:doaj.org-article:9e387ac290914899a72d3d48062a092f2021-12-02T11:39:20ZEvaluation of UBE3A antibodies in mice and human cerebral organoids10.1038/s41598-021-85923-x2045-2322https://doaj.org/article/9e387ac290914899a72d3d48062a092f2021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-85923-xhttps://doaj.org/toc/2045-2322Abstract UBE3A is an E3 ubiquitin ligase encoded by the neurally imprinted UBE3A gene. The abundance and subcellular distribution of UBE3A has been the topic of many previous studies as its dosage and localization has been linked to neurodevelopmental disorders including Autism, Dup15q syndrome, and Angelman syndrome. While commercially available antibodies have been widely employed to determine UBE3A localization, an extensive analysis and comparison of the performance of different UBE3A antibodies has not been conducted. Here we evaluated the specificities of seven commercial UBE3A antibodies in two of the major experimental models used in UBE3A research, mouse and human pluripotent stem cell-derived neural cells and tissues. We tested these antibodies in their two most common assays, immunofluorescence and western blot. In addition, we also assessed the ability of these antibodies to capture dynamic spatiotemporal changes of UBE3A by utilizing human cerebral organoid models. Our results reveal that among the seven antibodies tested, three antibodies demonstrated substantial nonspecific immunoreactivity. While four antibodies show specific localization patterns in both mouse brain sections and human cerebral organoids, these antibodies varied significantly in background signals and staining patterns in undifferentiated human pluripotent stem cells.Dilara SenZuzana DrobnaAlbert J. KeungNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Dilara Sen Zuzana Drobna Albert J. Keung Evaluation of UBE3A antibodies in mice and human cerebral organoids
description	Abstract UBE3A is an E3 ubiquitin ligase encoded by the neurally imprinted UBE3A gene. The abundance and subcellular distribution of UBE3A has been the topic of many previous studies as its dosage and localization has been linked to neurodevelopmental disorders including Autism, Dup15q syndrome, and Angelman syndrome. While commercially available antibodies have been widely employed to determine UBE3A localization, an extensive analysis and comparison of the performance of different UBE3A antibodies has not been conducted. Here we evaluated the specificities of seven commercial UBE3A antibodies in two of the major experimental models used in UBE3A research, mouse and human pluripotent stem cell-derived neural cells and tissues. We tested these antibodies in their two most common assays, immunofluorescence and western blot. In addition, we also assessed the ability of these antibodies to capture dynamic spatiotemporal changes of UBE3A by utilizing human cerebral organoid models. Our results reveal that among the seven antibodies tested, three antibodies demonstrated substantial nonspecific immunoreactivity. While four antibodies show specific localization patterns in both mouse brain sections and human cerebral organoids, these antibodies varied significantly in background signals and staining patterns in undifferentiated human pluripotent stem cells.
format	article
author	Dilara Sen Zuzana Drobna Albert J. Keung
author_facet	Dilara Sen Zuzana Drobna Albert J. Keung
author_sort	Dilara Sen
title	Evaluation of UBE3A antibodies in mice and human cerebral organoids
title_short	Evaluation of UBE3A antibodies in mice and human cerebral organoids
title_full	Evaluation of UBE3A antibodies in mice and human cerebral organoids
title_fullStr	Evaluation of UBE3A antibodies in mice and human cerebral organoids
title_full_unstemmed	Evaluation of UBE3A antibodies in mice and human cerebral organoids
title_sort	evaluation of ube3a antibodies in mice and human cerebral organoids
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/9e387ac290914899a72d3d48062a092f
work_keys_str_mv	AT dilarasen evaluationofube3aantibodiesinmiceandhumancerebralorganoids AT zuzanadrobna evaluationofube3aantibodiesinmiceandhumancerebralorganoids AT albertjkeung evaluationofube3aantibodiesinmiceandhumancerebralorganoids
_version_	1718395716299128832

Evaluation of UBE3A antibodies in mice and human cerebral organoids

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