Author: Avirutnan, Panisadee; Hauhart, Richard E.; Marovich, Mary A.; Garred, Peter; Atkinson, John P.; Diamond, Michael S.
Title: Complement-Mediated Neutralization of Dengue Virus Requires Mannose-Binding Lectin Document date: 2011_12_13
ID: 1x3n5job_34
Snippet: Hemolysis assay for serum C4. Serial dilutions of serum and purified C4 (Complement Technology) were prepared in DGVB Ï©Ï© (2.5 mM Bar-bital sodium, 139 mM dextrose, 71 mM NaCl, 0.1% [wt/vol] gelatin, 0.15 mM CaCl 2 , and 1 mM MgCl 2 ). Ten microliters of diluted serum or a C4 standard was added to a tube containing 476 l DGVB Ï©Ï© , 11 l of antibody-coated sheep erythrocytes (EA) (5 Ï« 10 8 cells/ml) (Complement Technology), and 3 l of C4-defici.....
Document: Hemolysis assay for serum C4. Serial dilutions of serum and purified C4 (Complement Technology) were prepared in DGVB ϩϩ (2.5 mM Bar-bital sodium, 139 mM dextrose, 71 mM NaCl, 0.1% [wt/vol] gelatin, 0.15 mM CaCl 2 , and 1 mM MgCl 2 ). Ten microliters of diluted serum or a C4 standard was added to a tube containing 476 l DGVB ϩϩ , 11 l of antibody-coated sheep erythrocytes (EA) (5 ϫ 10 8 cells/ml) (Complement Technology), and 3 l of C4-deficient guinea pig serum (Complement Technology). The tubes were incubated for 30 minutes at 37°C and then centrifuged for 5 minutes at 830 ϫ g. The OD of the supernatant was measured at 414 nm. C4 concentrations were calculated from a standard curve.
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