IgY Antibodies as Secondary Reagent in FACS Analysis

Avian vitellin antibodies (IgY) obtained after immunization with mouse IgG and mouse IgM in the presence or absence of Freund"s complete adjuvant are suited for labelling with FITC or biotin. In FACS analysis the antibodies prove to be excellent tools in detecting mouse monoclonal antibodies of different isotypes. The working dilution (titer of IgY-FITC) is highest when egg preparations of the secondary and tertiary immune response were used. Double staining of two different antigens can be carried out with direct labelled mab (FITC) and indirect labelled mab by IgY-conjugates (biotin). The two antigens can also be detected by double staining of different isotypes (mabs) by hen anti-mouse IgM FITC and hen anti-mouse IgG biotin-streptavidin-phycoerythrin. The staining obtained with IgY is at least as good or even better than that gained with mammalian secondary antibodies.