Human factor D of the alternative pathway: purification and quantitation by enzyme amplified electroimmunoassay.
Purified factor D was prepared with a yield of about 30%. Monospecific antiserum was raised in rabbits. Immunochemical quantitation of factor D in serum and plasma was performed by electroimmunoassay and a sensitive staining technique based on enzyme amplification. Peroxidase-labeled swine antibodies to rabbit immunoglobulin were applied to the gel after electrophoresis. Immune precipitates were then visualized by staining for peroxidase activity. Factor D could be detected at 50 micrograms/l. The normal concentration of D in serum was 1.6 mg/l, as assessed by this assay.
D000818:Animals; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D003166:Complement Activating Enzymes / Q000302:isolation & purification*; D011416:Complement Factor D / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D003378:Counterimmunoelectrophoresis / Q000379:methods*; D004492:Edetic Acid / Q000494:pharmacology; D006801:Humans; D007122:Immunoelectrophoresis / Q000379:methods*; D007124:Immunoenzyme Techniques; D011817:Rabbits
D004492:Edetic Acid; D003166:Complement Activating Enzymes; C493488:CFD protein, human; D011416:Complement Factor D