Journal: Veterinary Immunology and Immunopathology
Article Title: Quantification and phenotypic characterisation of peripheral IFN-γ producing leucocytes in chickens vaccinated against Newcastle disease
doi: 10.1016/j.vetimm.2017.10.001
Figure Lengend Snippet: Intracellular staining of IFN-γ in transfected CHO cells. A) CHO cells were either transfected with plasmids containing the chicken IFN-γ gene or empty plasmids (mock controls). Cells (day 22 after transfection) were cultured with or without BFA for 18 hours and subsequently stained intracellularly using monoclonal anti-chicken IFN-γ antibody (Mab80) conjugated with APC. Representative samples are show with frequency of IFN-γ+ cells indicated above gate. B) Comparison of transfected CHO cells (day 30 after transfection) cultured with 10 μg/ml BFA for 18 hours and subsequently stained intracellularly with either commercial ELISA capture antibody (5C.123.08) & A647 conjugated secondary anti-mouse IgG1 or monoclonal anti-chicken IFN-γ antibody directly conjugated with APC (Mab80) or commercial ELISA detection antibody (5C.123.02) directly conjugated with APC or polyclonal rabbit anti-chicken IFN-γ antibody & FITC-conjugated secondary goat anti-rabbit IgG. Representative samples are show with frequency of IFN-γ+ cells indicated above gate.
Article Snippet: Subsequently, the cells were washed twice with BD Perm/Wash buffer for 5 min at 600 × g , followed by staining with rabbit polyclonal anti-chIFN-γ IgG antibody (BioRad) and secondary goat anti-rabbit IgG FITC (Beckman Coulter, Brea, CA, cat no 732745).
Techniques: Staining, Transfection, Cell Culture, Comparison, Enzyme-linked Immunosorbent Assay