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Monoclonal Antibodies

Anti-IFN-γ [N2P2D5*A10]

Invented by Ayham Alnabulsi from Vertebrate Antibodies Limited
Invented at Vertebrate Antibodies Limited
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Production of four monoclonal antibodies against salmonid IFN-γ protein. Four distinct synthetic peptides containing 10 amino acid regions were selected to generate the antibodies.
The reactivity of four monoclonal antibodies generated against salmon and trout rIFN-γ was analysed by Western blotting. Results confirmed that all mAbs recognized the rIFN-γ (18 KDa), but not the trout rIL-21 (negative control):
Recognition of trout IFN-γ within head kidney cells stimulated with trout rIL-2 in vitro. Samples were lysed in Tri-Reagent and RNA and protein were extracted for analysis. Results from qPCR (A) and WB (B) were correlated, and 72-h incubation of HK cells with rIL-2 showed a high level of IFN-γ:
To verify the reactivity of produced mAbs against their specific peptides and the rIFN-γ an indirect-ELISA was developed. Results confirmed that mAbs recognised their specific peptides and the trout rIFN-γ but not trout rIL-21 (negative control):
To further characterise the produced mAbs, trout head kidney samples were subjected to 72-h incubation with PBS or trout rIL-2 and rIL-21. Cells were stained intracellularly with all IFN-γ mAbs and analysed in a BD AccuriTM C6. Preliminary results showed that V91 mAb recognises 5.8% fluorescent cells within the lymphocyte gate of non-stimulated HK cells. After 72h-stimulation with trout rIL-2 and rIL-21, this increased to 17.9%.
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Catalogue Number 152858
Applications ELISA FACS WB
Antigen/Gene or Protein Targets Interferon-gamma (IFNγ)
Synonyms IFG, IFI, IFN gamma, IFN, immune, IFN-gamma, IFNG, IFNG_HUMAN, Immune interferon, Interferon gamma
Reactivity Salmon and Rainbow Trout
Relevance Monoclonal antibody to detect IFNγ in fish models.

Background and Research Application
The zebrafish genome contains ten genes that encode class II cytokine-like peptides, of which the two that are related most closely to mammalian interferon gamma (IFN-gamma) were named IFN-γ1and IFN-γ2. IFNγ, or type II interferon, is a cytokine that is critical for innate and adaptive immunity against viral, some bacterial and protozoal infections. IFNγ is an important activator of macrophages and inducer of Class II major histocompatibility complex (MHC) molecule expression.
Host Mouse
Immunogen Peptide sequence - see figure
Immunogen UniProt ID Q5QRA1
Positive Control WB - recombinant IFNγ and kidney lysates, FACS - isolated leukocytes, kidney and spleen cells
Subclass Not Known
Myeloma Used P3X63Ag8.653
Strain Balb/c
Notes Production Details
Purified using multi-step affinity chromatography with protein A.

Storage Conditions
Store at -20 degrees frozen. Avoid repeated freeze/thaw cycles.

Concentration
1mg/ml as standard
Research Area Aquaculture

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