Identification of bluetongue virus protein-specific antibody responses in sheep by immunoblotting.

M. A. Adkison, J. L. Stott, B. I. Osburn

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Abstract

A western immunoblotting procedure was developed for identifying bluetongue virus protein-specific antibody responses in sheep. Assay conditions were optimized and included electrophoretic transfer of viral proteins to a nitrocellulose membrane (NCM), blocking of unbound sites on the NCM, and detection of NCM-bound primary immune complexes. A biotin-avidin-enzyme system was determined to be superior in terms of sensitivity for identification of the NCM-bound virus protein-antibody complexes. The biotin-avidin-enzyme detection system made use of biotinylated rabbit anti-sheep IgG and horseradish peroxidase-conjugated egg white avidin. This system permitted identification of antibodies specific for up to 10 bluetongue virus-associated proteins, 4 of which have not been characterized.

Original languageEnglish (US)
Pages (from-to)1194-1198
Number of pages5
JournalAmerican Journal of Veterinary Research
Volume48
Issue number8
StatePublished - Aug 1987

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ASJC Scopus subject areas

  • veterinary(all)

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