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Animals. The IgG fraction was isolated by use of protein G affinity chromatography d performed in accordance with the manufacturer's protocol; the IgG fraction then was biotinylated as described elsewhere. 17 The purified IgG was absorbed for 60

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in American Journal of Veterinary Research

. After blocking, membranes were incubated with ET-A or ET-B antisera (dilution, 1:500) for 1 hour at 25°C. Membranes were then washed with TBSS containing 0.1% Tween and incubated with donkey anti-sheep IgG−horseradish peroxidase conjugate. m After

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in American Journal of Veterinary Research

room temperature (approx 22°C) and with anti-desmin j (dilution, 1/200) or anti-SMMHC IgG k (dilution, 1/300) antibody for 1 and 2 hours, respectively, at room temperature. After several washes, cells were incubated with fluorescent dye

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in American Journal of Veterinary Research

, the most severe disease forms are observed in the young individuals; adults have milder clinical signs, and reinfection is common. In contrast to many viruses that primarily elicit IgG and cellular immune responses, RSV has the ability to skew the

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in American Journal of Veterinary Research

:250). Specificity of the immune serum was confirmed elsewhere. 5 Sections were again washed, and peroxide-conjugated goat anti-rabbit IgG p (1:400) was applied; sections then were incubated for 45 minutes at 21°C. Sections were washed, incubated (30 minutes at 21

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in American Journal of Veterinary Research

-Mouse IgG (H+L) Antibody, Molecular Probes, Eugene, Ore. e. Axiovert 200 M, Carl Zeiss, Oberkochen, Germany. f. Monoclonal Mouse Anti-Human Neurofilament Protein, Dako, Carpinteria, Calif. g. Polyclonal Goat Anti Mouse Immunoglobulin/Biotinylated, Dako

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in American Journal of Veterinary Research

were 24 hours old and used for an IgG enzyme immunoassay b to evaluate adequate transfer of passive immunity. Serial physical examinations and CBCs c were performed on days 5 and 8 to confirm health status of the foals. Physical examinations were

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in American Journal of Veterinary Research

confirm their health status prior to inclusion in the study. Adequate transfer of passive immunity was confirmed prior to initiation of the study by measurement of plasma IgG concentration via a commercial immunoassay. a The Institutional Animal Care and

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in American Journal of Veterinary Research

incubated overnight at 4°C with antibodies against tenascin-C (1:100), u TGF-β (1:100), v or VEGF (1:100). w To ensure antibody binding specificity, a peptide blocking v,w step was used for antibodies against TGF-β and VEGF, and nonspecific rabbit IgG

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in American Journal of Veterinary Research