Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Zhou, Xun, Shirokova, Vera, Kaminskyy, Vitaliy O., Berenger, Eva, Kochetkova, Elena, Norberg, Erik, Genander, Maria, Vakifahmetoglu-Norberg, Helin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2023
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
_version_ 1785083022491516928
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
work_keys_str_mv AT zhouxun knockoutvalidationoflamp2aantibodiesforimmunostaininginhumancancercells
AT shirokovavera knockoutvalidationoflamp2aantibodiesforimmunostaininginhumancancercells
AT kaminskyyvitaliyo knockoutvalidationoflamp2aantibodiesforimmunostaininginhumancancercells
AT berengereva knockoutvalidationoflamp2aantibodiesforimmunostaininginhumancancercells
AT kochetkovaelena knockoutvalidationoflamp2aantibodiesforimmunostaininginhumancancercells
AT norbergerik knockoutvalidationoflamp2aantibodiesforimmunostaininginhumancancercells
AT genandermaria knockoutvalidationoflamp2aantibodiesforimmunostaininginhumancancercells
AT vakifahmetoglunorberghelin knockoutvalidationoflamp2aantibodiesforimmunostaininginhumancancercells