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determine the HAI titers in infected mice, sera of these were treated with receptor-destroying enzyme (RDE/Cholera filtrate; Sigma) to remove sialic acids from serum proteins capable of aspecific inhibition of agglutination. After incubation overnight at 37°C, the RDE was inactivated by addition of 0.75% sodium citrate in PBS and heating to 56°C for 30 min. To remove sialic acid binding proteins, sera were cleared with 1/10 volume 50% chicken RBC. Titration was done by incubating a two-fold dilution series of sera with 4 HA units of X-47 virus for 1 hour at room temperature in 96-well U-bottom plates. Finally, an equal volume of 0.5% chicken RBC was added and titers were read 30 min later. Negative controls included PBS instead of immune serum (agglutination control) or PR8 instead of X-47 virus (control for agglutination effect of sera); as positive control, serum from a mouse infected twice with a sublethal dose of X-47 was used."}

{"project":"UBERON-AE","denotations":[{"id":"T15659","span":{"begin":875,"end":880},"obj":"http://purl.obolibrary.org/obo/UBERON_0001977"},{"id":"T15658","span":{"begin":751,"end":756},"obj":"http://purl.obolibrary.org/obo/UBERON_0001977"},{"id":"T15657","span":{"begin":163,"end":168},"obj":"http://purl.obolibrary.org/obo/UBERON_0001977"}],"text":"To determine the HAI titers in infected mice, sera of these were treated with receptor-destroying enzyme (RDE/Cholera filtrate; Sigma) to remove sialic acids from serum proteins capable of aspecific inhibition of agglutination. After incubation overnight at 37°C, the RDE was inactivated by addition of 0.75% sodium citrate in PBS and heating to 56°C for 30 min. To remove sialic acid binding proteins, sera were cleared with 1/10 volume 50% chicken RBC. Titration was done by incubating a two-fold dilution series of sera with 4 HA units of X-47 virus for 1 hour at room temperature in 96-well U-bottom plates. Finally, an equal volume of 0.5% chicken RBC was added and titers were read 30 min later. Negative controls included PBS instead of immune serum (agglutination control) or PR8 instead of X-47 virus (control for agglutination effect of sera); as positive control, serum from a mouse infected twice with a sublethal dose of X-47 was used."}

{"project":"GO-BP","denotations":[{"id":"T15672","span":{"begin":823,"end":836},"obj":"http://purl.obolibrary.org/obo/GO_0007157"},{"id":"T15671","span":{"begin":758,"end":771},"obj":"http://purl.obolibrary.org/obo/GO_0007157"},{"id":"T15670","span":{"begin":213,"end":226},"obj":"http://purl.obolibrary.org/obo/GO_0007157"},{"id":"T15669","span":{"begin":823,"end":836},"obj":"http://purl.obolibrary.org/obo/GO_0000752"},{"id":"T15668","span":{"begin":758,"end":771},"obj":"http://purl.obolibrary.org/obo/GO_0000752"},{"id":"T15667","span":{"begin":213,"end":226},"obj":"http://purl.obolibrary.org/obo/GO_0000752"}],"text":"To determine the HAI titers in infected mice, sera of these were treated with receptor-destroying enzyme (RDE/Cholera filtrate; Sigma) to remove sialic acids from serum proteins capable of aspecific inhibition of agglutination. After incubation overnight at 37°C, the RDE was inactivated by addition of 0.75% sodium citrate in PBS and heating to 56°C for 30 min. To remove sialic acid binding proteins, sera were cleared with 1/10 volume 50% chicken RBC. Titration was done by incubating a two-fold dilution series of sera with 4 HA units of X-47 virus for 1 hour at room temperature in 96-well U-bottom plates. Finally, an equal volume of 0.5% chicken RBC was added and titers were read 30 min later. Negative controls included PBS instead of immune serum (agglutination control) or PR8 instead of X-47 virus (control for agglutination effect of sera); as positive control, serum from a mouse infected twice with a sublethal dose of X-47 was used."}

{"project":"GO-MF","denotations":[{"id":"T15675","span":{"begin":385,"end":392},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T15674","span":{"begin":385,"end":401},"obj":"http://purl.obolibrary.org/obo/GO_0005515"},{"id":"T15673","span":{"begin":373,"end":392},"obj":"http://purl.obolibrary.org/obo/GO_0033691"}],"text":"To determine the HAI titers in infected mice, sera of these were treated with receptor-destroying enzyme (RDE/Cholera filtrate; Sigma) to remove sialic acids from serum proteins capable of aspecific inhibition of agglutination. After incubation overnight at 37°C, the RDE was inactivated by addition of 0.75% sodium citrate in PBS and heating to 56°C for 30 min. To remove sialic acid binding proteins, sera were cleared with 1/10 volume 50% chicken RBC. Titration was done by incubating a two-fold dilution series of sera with 4 HA units of X-47 virus for 1 hour at room temperature in 96-well U-bottom plates. Finally, an equal volume of 0.5% chicken RBC was added and titers were read 30 min later. Negative controls included PBS instead of immune serum (agglutination control) or PR8 instead of X-47 virus (control for agglutination effect of sera); as positive control, serum from a mouse infected twice with a sublethal dose of X-47 was used."}

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