Encoded by the C6 gene in humans, complement component 6 (C6) is the protein involved in the complement system. This component can cause cell lysis by intercalating into the cell membrane, so C6 deficiency results in greater susceptibility to bacterial infection. As a professional product and service provider in the field of complement, Creative Biolabs has developed a series of methods and protocols. Here, we describe the C6 purification protocol for our clients all over the world.
C6 Purification
Fig.1 Flow chart of C6 purification protocol. (Creative Biolabs)
Published Data
The complement membrane attack complex (MAC) forms sequentially through the assembly of C5b with homologous proteins-comprising a single copy each of C6, C7, and C8, in addition to 12-14 molecules of C9-to construct a lytic pore within bacterial membranes. The MAC-perforin domain, composed of C6 through C9, is flanked by several auxiliary domains. Researchers purified complement C6 from human plasma using an adapted protocol. This method involved barium citrate depletion of vitamin K-dependent proteins, precipitation with 4–12% polyethylene glycol, DEAE-Sephadex chromatography, euglobulin precipitation, gel filtration on Sephacryl-300, dextran sulfate-Sepharose chromatography, and IgG-Sepharose 6B antibody purification, thereby isolating the first and longest pore protein recruited by C5b.
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