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Monoclonal antibodies isolated by large-scale screening are suitable for labeling adult zebrafish (Danio rerio) tissues and cell structures.
- Source :
-
Journal of immunological methods [J Immunol Methods] 2009 Jul 31; Vol. 346 (1-2), pp. 9-17. Date of Electronic Publication: 2009 May 03. - Publication Year :
- 2009
-
Abstract
- The lack of a sufficient number of antibodies represents an obstacle in the research performed using the zebrafish (Danio rerio) as a model organism. On the other hand, high-throughput generation of antibodies, especially those suitable for immunohistochemistry, is not an established methodology. Here we present the results of an immunization experiment with a zebrafish tissue lysate that allowed for the isolation of hundreds of monoclonal antibodies suitable for labeling of a large variety of zebrafish tissue and cell structures. Some of them were further characterized in terms of detailed localization and age-dependent expression. In addition, the antigen recognized by one of them was first immunoprecipitated and then identified by mass spectrometry. Furthermore, immunofluorescence-competent recombinant antibodies were also isolated by panning large repertoire phage display libraries, in both single-chain (scFv) and single-domain (VHH) format. Such selection alternative is simpler to organize and could contribute to limit the costs of antibody screening and production.
- Subjects :
- Age Factors
Animals
Antibodies, Monoclonal biosynthesis
Antibodies, Monoclonal genetics
Blotting, Western
Epitope Mapping
Hybridomas metabolism
Immunoglobulin Heavy Chains genetics
Immunoglobulin Heavy Chains isolation & purification
Immunoglobulin Variable Region genetics
Immunoglobulin Variable Region isolation & purification
Immunohistochemistry
Immunoprecipitation
Mice
Mice, Inbred BALB C
Microscopy, Fluorescence
Recombinant Proteins isolation & purification
Zebrafish
Zebrafish Proteins analysis
Antibodies, Monoclonal isolation & purification
Immunization
Peptide Library
Zebrafish Proteins immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1872-7905
- Volume :
- 346
- Issue :
- 1-2
- Database :
- MEDLINE
- Journal :
- Journal of immunological methods
- Publication Type :
- Academic Journal
- Accession number :
- 19410577
- Full Text :
- https://doi.org/10.1016/j.jim.2009.04.012