Journal: Animals : an Open Access Journal from MDPI

Article Title: Development of a Monoclonal Antibody Against Duck IFN-γ Protein and the Application for Intracellular Cytokine Staining

doi: 10.3390/ani15060815

Figure Lengend Snippet: Characterization of mAb against duck IFN-γ protein. ( A ) Morphologic observation of 24H4 hybridoma cell. Scale bar, 100 μm. ( B ) Western blot identification of mAb. Lane M: protein molecular weight standard; Lane 1: Purified duIFN-γ-Fc protein. The purified duIFN-γ mAb was used as the primary antibody, and the goat anti-mouse IgG antibody was used as the secondary antibody. ( C ) IFN-γ antibody sensitivity was determined by indirect ELISA using duIFN-γ-Fc protein as the antigen and mAb 24H4 as the primary antibody. S/P ratio = sample OD 450 /NC OD 450 , S/P > 2 was considered positive. ( D ) Subclass of the mAb 24H4 was identified using a mouse antibody homotype ELISA kit; **** on the bar represents an extremely significant difference ( p < 0.0001).

Article Snippet: The cells were then stained with FITC-conjugated Goat Anti-Mouse IgG2b (SouthernBiotech, Birmingham, UK).

Techniques: Western Blot, Molecular Weight, Purification, Indirect ELISA, Enzyme-linked Immunosorbent Assay