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Int. J. Mol. Sci. 2018, 19(1), 14; doi:10.3390/ijms19010014

Generation and Characterization of Antibodies against Opioid Receptors from Zebrafish

1
Department of Cell Biology and Pathology, Instituto de Neurociencias de Castilla y León (INCyL), Universidad de Salamanca, 37007 Salamanca, Spain
2
Institute of Biomedical Research of Salamanca (IBSAL), University Hospital of Salamanca, 37007 Salamanca, Spain
3
Department of Biochemistry and Molecular Biology, Instituto de Neurociencias de Castilla y León (INCyL), Universidad de Salamanca, 37007 Salamanca, Spain
Current address: Tumor Immunology Laboratory, IdiPAZ, La Paz Hospital, Paseo de la Castellana 261, 28406 Madrid, Spain.
Current address: Department of Chemistry and Biochemistry, University of Colorado, Boulder, CO 80309, USA.
§
Current address: Department of Genetics, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA.
*
Author to whom correspondence should be addressed.
Received: 14 November 2017 / Revised: 4 December 2017 / Accepted: 8 December 2017 / Published: 2 January 2018
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
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Abstract

The opioid system is well conserved among species and plays a critical role in pain and addiction systems. The use of zebrafish as an experimental model to study development and genetics is extraordinary and has been proven to be relevant for the study of different diseases. The main drawback to its use for the analysis of different pathologies is the lack of protein tools. Antibodies that work in other models are not suitable for zebrafish due to the low degree of homology that exists among the opioid receptor protein sequences in different species. Here we report the successful generation and characterization of antibodies against the mu, delta 1 and delta 2 opioid receptors in zebrafish. The antibodies obtained, which are specific for each receptor due to the use of the C-terminus as antigens, work for Western blotting and immunohistochemistry. In addition, the antibodies against mu and delta 1 opioid receptors, but not those against delta 2, are able to immunoprecipitate the corresponding receptor from zebrafish lysates. The development of opioid receptor antibodies is an asset to the further study of the endogenous opioid system in zebrafish. View Full-Text
Keywords: opioid receptors; antibodies; zebrafish opioid receptors; antibodies; zebrafish
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Arévalo, J.C.; Hernández-Jiménez, E.; Jiménez-González, A.; Torres-Valle, M.; Iwasaki, R.S.; López-Bellido, R.; Vicente-García, C.; Rodríguez, R.E. Generation and Characterization of Antibodies against Opioid Receptors from Zebrafish. Int. J. Mol. Sci. 2018, 19, 14.

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