ALTEX :: Alternatives to Animal Experiments

1996, Supplement

IgY Antibodies as Secondary Reagent in FACS Analysis

Undine Hommel, Ingrid Behn and Sunna Hauschildt

University of Leipzig, Institute of Zoology/Immunobiology, D-Leipzig

SUMMARY

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 labeled mab (FITC) and indirect labeled 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. (article in english)

Keywords: Avian vitellin antibodies, IgY, labeling, FACS analysis