Figure 2

A polyclonal anti-protein C antibody specifically detecting CnPC in plasma. (A) Silver stained reducing 12% SDS-PAGE of human protein C-deficient plasma and with added CnPC. Lane 1: CnPC (0.5 μg) purified from canine plasma. Lane 2: Human protein C-deficient plasma (5 μg of protein). Lane 3: Human protein C-deficient plasma (5 μg of protein) spiked with 0.5 μg of CnPC. Lane 4: Molecular weight markers. (B) Western blot of gel in (A) probed with a polyclonal antibody to EDQVDPRLIDGK. Lane loads were the same as (A). Two tightly spaced bands (likely two glycoforms) were present in lanes 1 and 3, indicating detection of CnPC. Only the major glycoform was visible in lane 1 in (A), and this was likely related to lower sensitivity of silver staining than western immunoblotting.