Preparation and Evaluation of Anti Rota IgG Conjugated with Fluorescein
Preparation and Evaluation of Anti Rota IgG Conjugated with Fluorescein
Arwa H. El -Naggar, Nirmeen G. Shafiek, and M. S. Wassel.
ABSTRACT
Hyper immune serum against bovine Rota was prepared in guinea pigs through its inoculation with complete and incomplete Freund's adjuvant and without adjuvant in successive doses into susceptible guinea pigs. Serum neutralization (SNT), ELISA and indirect fluorescent assay (IFA) tests were used to evaluate the hyper immune serum Before conjugation with fluorescent. The antibody titer reach 32 by SNT and 5984 by ELISA and identified by IFA to be Bovine rota. Separation of anti B. Rota immunoglobulins IgG "were done using activated Sepharose 4B followed by conjugation with fluorescein isothiocyanate at PH 9.6. The conjugated IgG was tested against reference B. Rota antigen using direct and indirect fluorescein assay (FA. IFA). Both assay gave good result as the working dilution of the fluorescein isothiocyanate conjugated B.rota IgG was 1:30.
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