4613347777

Enzyme-Iinked immunosorbent assay

C3 molecule concentration was tested in mice sera at the indicated time points by EL1SA. Briefly, blood was harvested and incubated at 37°C to coagulate. Samples were centrifuged and the serum fraction was used. Serial dilutions were added onto an IgM-coated 96-well piąte (5pg/ml, Sigma-Aldrich, Oakville, ON). Mouse anti-C3 coupled to biotin (Cedarlane, Burlington, ON) followed with streptavidin-horseradish peroxidase (Southern Biotech, Birmingham, AL) were incubated with samples. A solution of o-Phenylenediamine dihydrochloride (Sigma-Aldrich, Oakville, ON) was then used for the detection of C3 molecules. Optical density was measured by spectrophotometry at 490 nm.