Fit-for-purpose based testing and validation of antibodies to amino- and carboxy-terminal domains of cannabinoid receptor 1
dc.contributor.author | Echeazarra Escudero, Leire | |
dc.contributor.author | García del Caño, Gontzal | |
dc.contributor.author | Barrondo Lacarra, Sergio ![]() | |
dc.contributor.author | González Burguera, Imanol | |
dc.contributor.author | Saumell Esnaola, Miquel | |
dc.contributor.author | Aretxabala Rodríguez, Xabier | |
dc.contributor.author | López de Jesús, Maider ![]() | |
dc.contributor.author | Borrega Román, Leire | |
dc.contributor.author | Mato Santos, Susana ![]() | |
dc.contributor.author | Ledent, Catherine | |
dc.contributor.author | Matute Almau, Carlos José | |
dc.contributor.author | Goicolea Altuna, María Aranzazu ![]() | |
dc.contributor.author | Sallés Alvira, Joan | |
dc.date.accessioned | 2022-01-20T13:10:47Z | |
dc.date.available | 2022-01-20T13:10:47Z | |
dc.date.issued | 2021-11 | |
dc.identifier.citation | Histochemistry and Cell Biology 156(5) : 479-502 (2021) | es_ES |
dc.identifier.issn | 0948-6143 | |
dc.identifier.issn | 1432-119X | |
dc.identifier.uri | http://hdl.handle.net/10810/55073 | |
dc.description.abstract | Specific and selective anti-CB1 antibodies are among the most powerful research tools to unravel the complex biological processes mediated by the CB1 receptor in both physiological and pathological conditions. However, low performance of antibodies remains a major source of inconsistency between results from different laboratories. Using a variety of techniques, including some of the most commonly accepted ones for antibody specificity testing, we identified three of five commercial antibodies against different regions of CB1 receptor as the best choice for specific end-use purposes. Specifically, an antibody against a long fragment of the extracellular amino tail of CB1 receptor (but not one against a short sequence of the extreme amino-terminus) detected strong surface staining when applied to live cells, whereas two different antibodies against an identical fragment of the extreme carboxy-terminus of CB1 receptor (but not one against an upstream peptide) showed acceptable performance on all platforms, although they behaved differently in immunohistochemical assays depending on the tissue fixation procedure used and showed different specificity in Western blot assays, which made each of them particularly suitable for one of those techniques. Our results provide a framework to interpret past and future results derived from the use of different anti-CB1 antibodies in the context of current knowledge about the CB1 receptor at the molecular level, and highlight the need for an adequate validation for specific purposes, not only before antibodies are placed on the market, but also before the decision to discontinue them is made. | es_ES |
dc.description.sponsorship | Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. This work was funded by Basque Government (IT1230-19, to J.S.), MINECO CTQ2017-85686-R (Spanish Ministry of Economy and Competitiveness, to J.S. and M.A.G.), FEDER and ISCIII (AES 2018-PI18/00513, to S.M.), MINECO SAF2016-75292-R MINECO, to C.M.) and Centro de Investigacion Biomedica en Red de Salud Mental (CIBERSAM, to J.S. and S.B.). | es_ES |
dc.language.iso | eng | es_ES |
dc.publisher | Springer | es_ES |
dc.relation | info:eu-repo/grantAgreement/MINECO/CTQ2017-85686-R | es_ES |
dc.relation | info:eu-repo/grantAgreement/MINECO/SAF2016-75292-R | es_ES |
dc.rights | info:eu-repo/semantics/openAccess | es_ES |
dc.rights.uri | http://creativecommons.org/licenses/by/3.0/es/ | * |
dc.subject | antibody specificity | es_ES |
dc.subject | CB1 receptor | es_ES |
dc.subject | carboxy-terminus | es_ES |
dc.subject | amino-terminus | es_ES |
dc.subject | antigen retrieval | es_ES |
dc.subject | CB1-knockout mice | es_ES |
dc.subject | phospholipase-C-beta | es_ES |
dc.subject | diacylglycerol lipase-alpha | es_ES |
dc.subject | protein-coupled receptors | es_ES |
dc.subject | acid amide hydrolase | es_ES |
dc.subject | CB1 receptor | es_ES |
dc.subject | endocannabinoid system | es_ES |
dc.subject | rat-brain | es_ES |
dc.subject | subcellular-localization | es_ES |
dc.subject | muscarinic acetylcholine | es_ES |
dc.subject | messenger-RNA | es_ES |
dc.title | Fit-for-purpose based testing and validation of antibodies to amino- and carboxy-terminal domains of cannabinoid receptor 1 | es_ES |
dc.type | info:eu-repo/semantics/article | es_ES |
dc.rights.holder | This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. | es_ES |
dc.rights.holder | Atribución 3.0 España | * |
dc.relation.publisherversion | https://link-springer-com.ehu.idm.oclc.org/article/10.1007%2Fs00418-021-02025-5#rightslink | es_ES |
dc.identifier.doi | 10.1007/s00418-021-02025-5 | |
dc.departamentoes | Farmacología | es_ES |
dc.departamentoes | Fisiología | es_ES |
dc.departamentoes | Neurociencias | es_ES |
dc.departamentoes | Química analítica | es_ES |
dc.departamentoeu | Farmakologia | es_ES |
dc.departamentoeu | Fisiologia | es_ES |
dc.departamentoeu | Kimika analitikoa | es_ES |
dc.departamentoeu | Neurozientziak | es_ES |
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