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Monoclonal Antibodies

Anti-IFN-γ [VAB3P2D2*C11]

Invented by Ayham Alnabulsi from Vertebrate Antibodies Limited
Invented at Vertebrate Antibodies Limited
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Image thumbnail for Anti-IFN-γ [VAB3P2D2*C11]
Image thumbnail for Anti-IFN-γ [VAB3P2D2*C11]
Image thumbnail for Anti-IFN-γ [VAB3P2D2*C11]
Image thumbnail for Anti-IFN-γ [VAB3P2D2*C11]
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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%.
Immunohistochemistry is showing IFN-gamma positive cells using Anti-IFN-γ [VAB3P2D2*C11] in PKD infected rainbow trout kidney. IHC was performed on formalin-fixed, paraffin-embedded tissue sections. Antigen retrieval step is required (antigen retrieval: microwave 20 min @ 800W in 10 mM citrate buffer, pH 6.0).
Immunohistochemistry image showing IFN-gamma positive cells using Anti-IFN-γ [VAB3P2D2*C11] in vaccinated rainbow trout head kidney. IHC was performed on formalin-fixed, paraffin-embedded tissue sections. Antigen retrieval step is required (antigen retrieval: microwave 20 min @ 800W in 10 mM citrate buffer, pH 6.0).
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Catalogue Number 152861
Applications ELISA FACS IHC 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 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-gamma1 and IFN-gamma2.
Host Mouse
Immunogen Peptide sequence - see figure
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
Research Area Aquaculture

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