Format

Send to

Choose Destination
Fish Shellfish Immunol. 2016 Oct;57:79-86. doi: 10.1016/j.fsi.2016.08.023. Epub 2016 Aug 15.

Analysis of interferon gamma protein expression in zebrafish (Danio rerio).

Author information

1
Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, UK. Electronic address: sohyeyoon@abdn.ac.uk.
2
Vertebrate Antibodies Ltd, Aberdeen, UK.
3
Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, UK.
4
Laboratory of Molecular Genetics, College of Bioscience and Biotechnology, National Cheng Kung University, Taiwan.
5
Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, UK; Molecular Genetics, Department of Biological Sciences, University of Waikato, Hamilton, New Zealand.
6
Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, UK. Electronic address: c.secombes@abdn.ac.uk.

Abstract

IFN-γ is a major effector cytokine, produced to induce type I immune responses. It has been cloned in several fish species including zebrafish, however to date few studies have looked at IFN-γ protein expression and bioactivity in fish. Hence, the current study focused on developing a monoclonal antibody (moAb) against zfIFN-γ. We show that the zfIFN-γ moAb specifically recognises E. coli produced recombinant IFN-γ protein and zfIFN-γ produced in transfected HEK293 cells, by Western blot analysis. Next we analysed the production of the native protein following expression induced by PHA stimulation of leukocytes in vitro or antigen re-stimulation in vivo. We show the IFN-γ protein is produced as a dimer, and that a good correlation exists between transcript expression levels and protein levels.

KEYWORDS:

Antigen re-stimulation; IFN-γ; Monoclonal antibody; PHA; Protein expression; Zebrafish

PMID:
27539703
DOI:
10.1016/j.fsi.2016.08.023
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center