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Knockout validation of LAMP2A antibodies for immunostaining in human cancer cells
LAMP2A is the rate-limiting factor of chaperone-mediated autophagy (CMA), a unique selective protein degradative pathway. To date LAMP2A antibodies are not knockout (KO)-validated in human cells. We have recently generated human isoform-specific LAMP2A KO cells, and here we assessed the specificity...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Taylor & Francis
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10392744/ https://www.ncbi.nlm.nih.gov/pubmed/37194363 http://dx.doi.org/10.1080/15548627.2023.2213515 |
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author | Zhou, Xun Shirokova, Vera Kaminskyy, Vitaliy O. Berenger, Eva Kochetkova, Elena Norberg, Erik Genander, Maria Vakifahmetoglu-Norberg, Helin |
author_facet | Zhou, Xun Shirokova, Vera Kaminskyy, Vitaliy O. Berenger, Eva Kochetkova, Elena Norberg, Erik Genander, Maria Vakifahmetoglu-Norberg, Helin |
author_sort | Zhou, Xun |
collection | PubMed |
description | LAMP2A is the rate-limiting factor of chaperone-mediated autophagy (CMA), a unique selective protein degradative pathway. To date LAMP2A antibodies are not knockout (KO)-validated in human cells. We have recently generated human isoform-specific LAMP2A KO cells, and here we assessed the specificity of select commercial LAMP2A antibodies on wild-type and LAMP2A KO human cancer cells. While all tested antibodies were suitable for immunoblotting, the anti-LAMP2A antibody (ab18528) is likely to exhibit an off-target reactivity in immunostaining approaches using human cancer cells, and alternative antibodies, which seem more appropriate, are available. |
format | Online Article Text |
id | pubmed-10392744 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Taylor & Francis |
record_format | MEDLINE/PubMed |
spelling | pubmed-103927442023-08-02 Knockout validation of LAMP2A antibodies for immunostaining in human cancer cells Zhou, Xun Shirokova, Vera Kaminskyy, Vitaliy O. Berenger, Eva Kochetkova, Elena Norberg, Erik Genander, Maria Vakifahmetoglu-Norberg, Helin Autophagy Resource LAMP2A is the rate-limiting factor of chaperone-mediated autophagy (CMA), a unique selective protein degradative pathway. To date LAMP2A antibodies are not knockout (KO)-validated in human cells. We have recently generated human isoform-specific LAMP2A KO cells, and here we assessed the specificity of select commercial LAMP2A antibodies on wild-type and LAMP2A KO human cancer cells. While all tested antibodies were suitable for immunoblotting, the anti-LAMP2A antibody (ab18528) is likely to exhibit an off-target reactivity in immunostaining approaches using human cancer cells, and alternative antibodies, which seem more appropriate, are available. Taylor & Francis 2023-05-16 /pmc/articles/PMC10392744/ /pubmed/37194363 http://dx.doi.org/10.1080/15548627.2023.2213515 Text en © 2023 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) ), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way. The terms on which this article has been published allow the posting of the Accepted Manuscript in a repository by the author(s) or with their consent. |
spellingShingle | Resource Zhou, Xun Shirokova, Vera Kaminskyy, Vitaliy O. Berenger, Eva Kochetkova, Elena Norberg, Erik Genander, Maria Vakifahmetoglu-Norberg, Helin Knockout validation of LAMP2A antibodies for immunostaining in human cancer cells |
title | Knockout validation of LAMP2A antibodies for immunostaining in human cancer cells |
title_full | Knockout validation of LAMP2A antibodies for immunostaining in human cancer cells |
title_fullStr | Knockout validation of LAMP2A antibodies for immunostaining in human cancer cells |
title_full_unstemmed | Knockout validation of LAMP2A antibodies for immunostaining in human cancer cells |
title_short | Knockout validation of LAMP2A antibodies for immunostaining in human cancer cells |
title_sort | knockout validation of lamp2a antibodies for immunostaining in human cancer cells |
topic | Resource |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10392744/ https://www.ncbi.nlm.nih.gov/pubmed/37194363 http://dx.doi.org/10.1080/15548627.2023.2213515 |
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