Figure 3.

Carboxymethyl cellulose-coated carbon nanotubes (CMC-CNTs) stably bind to purified human properdin and recombinant TSR4+5, as shown via SDS-PAGE (A) and western blot (B). (A) CMC-CNTs were incubated with recombinant human properdin, or thrombospondin type I repeat (TSR) 4 + 5-MBP fusion protein and BSA overnight in the affinity buffer. Carbon nanotubes (CNTs) were washed extensively via centrifugation and run on an SDS-PAGE (12%) under reduced conditions. Properdin migrated at ~55 kDa and MBP-TSR4+5 migrated as a single band at ~53 kDa. (B) In parallel, the SDS-PAGE was transferred onto nitrocellulose membrane for 2 h at 320 mA. The blot was probed with anti-human properdin (polyclonal) antibodies, followed by protein A–HRP conjugate and developed using 3,3′-diaminobenzidine.