Mouse anti his antiserum

Human glu-plasminogen was obtained from Haematologic Technologies (Essex Junction, VT, USA). Plasminogen was activated to plasmin using urokinase plasminogen activator (uPA) from Merck Millipore, Darmstadt, Germany. Both the chromogenic substrate S-2251 (D-Val-Leu-Lys p-nitroanilide dihydrochloride) and fibrinogen were purchased from Sigma-Aldrich (Steinheim, Germany). Goat anti-fibrinogen antiserum was purchased from Acris Antibodies (Herford, Germany), mouse anti-His antiserum was obtained from GE Healthcare (Munich, Germany). Horseradish peroxidase (HRP)-conjugated immunoglobulins were purchased from Dako (Hamburg, Germany).

Non-immune human serum (NHS) collected from healthy blood donors was initially tested for the presence of anti-Borrelia IgM and IgG antibodies as previously described (29 (link)). Only sera considered to be negative were combined to form a serum pool. All complement components were purchased from Complement Technology (Tyler, TX, USA). Polyclonal anti-FH and anti-C3 antisera were obtained from Merck Biosciences (Bad Soden, Germany) and the neoepitope-specific monoclonal anti-C5b-9 antibody was purchased from Quidel (San Diego, CA, USA). Purification of FHL-1, FH fragments containing different CCP domains, and the anti-CCP1-4 antiserum has been described previously (39 (link)). The monoclonal antibody L41 1C11 was used to detect the borrelial FlaB protein (40 (link)). Proteinase K and trypsin were purchased from Merck (Darmstadt, Germany). For the detection of His-tagged proteins, a mouse anti-His antiserum was used (GE Healthcare, Munich, Germany), and all horseradish peroxidase-conjugated immunoglobulins were purchased from Dako (Hamburg, Germany).