Immunoblotting of kininogen in murine plasma. (A) An antibody raised against the mHK1-specific sequence ERDAETEQGPTH was used to blot the in vitro–translated polypeptides encoded by the cloned PCR products derived from liver and kidney cDNA using the primers depicted in Figure 3B. In the top panel, these recombinant polypeptides were separated using 10% SDS-PAGE, and the gel was stained with Coomassie brilliant blue. In the bottom panel, the polypeptides were transferred to PVDF and immunoblotted with the mHK1-specific antipeptide antibody. Only the liver (mKng1)–derived polypeptide is recognized by this antibody, demonstrating specificity. (B) The affinity-purified anti-mHK1 antibody was used to blot plasma from wild-type (+/+) and 4 mKng1-deficient (−/−) mice; mHK1 was present only in the wild-type animals. (C) Immunoblotting of whole mouse plasma (1:10 and 1:20 dilutions) from WT and mKng1^−/− mice with an anti-BK antisera. A total of 3 distinct kininogen species are present in plasma from the wild-type mouse, but none are present in the mKng1^−/− mice. The upper band recognized by the antisera corresponds to mHK, the intermediate band corresponds to ΔmHK-D5, and the lower band corresponds to mLK.