Mouse Plasma Kallikrein/KLKB1 Antibody

Detection of Plasma Kallikrein/KLKB1 by Western Blot Klebsiella pneumoniae does not activate the contact system in vivo or in vitro. Mice were treated subcutaneously with PKK ASO (open bars) or control ASO (grey bars) twice weekly for 3 weeks; 4 days after final dose, mice were infected with K. pneumoniae via airways (t = 0) and euthanized at 12 or 36 h after infection. (A) Western blots showing full length PKK and the cleaved chain of alpha ‐PKa in plasma obtained 36 h after infection using either reduced or non‐reduced samples; naive mouse plasmas pretreated with 100 μg/ml kaolin for the indicated time (15 min to 1 h) were used as positive controls. (B, C) K. pneumoniae, E. coli Ymel or E. coli Ymel‐1 was incubated in citrated plasma from healthy volunteers and bacterial bound PKK (B) and HK (C) proteins were detected by western blotting after reducing SDS‐PAGE. cHK, cleaved chains of HK; alpha ‐PKahc, alpha ‐PKa heavy chain. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31595971), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Plasma Kallikrein/KLKB1 by Western Blot Inhibition of PKK expression by PKK ASO treatment. Mice were treated subcutaneously with PKK ASO (open bars) or control (Ctrl) ASO (grey bars) twice weekly for 3 weeks; 4 days after final dose, mice were infected with K. pneumoniae via the airways (t = 0). (A) Western blot of mouse plasma PKK (t = 0). Plasma (0.2 μl diluted in buffer; total volume 10 μl) from four PKK ASO‐ or four Ctrl ASO‐treated mice was loaded; ‘standard’ indicates volumes (μl) of plasma in buffer (total volume 10 μl) from one uninfected mouse (treated with Ctrl ASO). The blot shown is representative of three independent blots. (B) Klkb1 mRNA expression in livers of mice treated with PKK or Ctrl ASO at corresponding time points. (C) Plasma PKK activity. (D) Activated partial thromboplastin time (aPTT). (E) Prothrombin time (PT). Data are mean and SD for bar charts in B and C or medians for box and whisker plots in D and E, eight mice per group. **p < 0.01, ***p < 0.001 (versus Ctrl ASO at the same time point); φp < 0.05, φφp < 0.01, φφφp < 0.001 (versus t = 0 in the same treatment group). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31595971), licensed under a CC-BY license. Not internally tested by R&D Systems.