Purification and Characterization of Human and Mouse Complement C3

Complement component C3 is the most abundant complement protein in plasma, central to all three complement activation pathways and essential to complement amplification. Thus, it is one of the most extensively studied complement proteins. This chapter describes the purification of C3 from human and mouse plasma using protein precipitation, followed by classical ion exchange chromatography and gel filtration. The biochemical and functional characteristics of the purified C3 are typically assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and hemolysis assays. The hemolysis assay is a standard technique to assess complement activity monitoring the lysis of red blood cells.