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Fig. 2 | Molecular Medicine

Fig. 2

From: Mechanism of Action of Anti-C1-Inhibitor Autoantibodies: Prevention of the Formation of Stable C1s-C1-inh Complexes

Fig. 2

The effect of synthetic peptides on anti-C1-inh autoantibodies. The concentrations of C1-inh, 125I-labeled C1s, and autoantibodies used in the reaction were 2.1 µM, 1.8 µM (75,000 cpm), and 0.5 µM, respectively. The concentration of the autoantibody used was calculated to cause about 50% inhibition of complex formation (see Fig. 5A). All protein reagents were diluted with PBS-1% Triton ×100 and the reaction volume was set to a total volume of 40 /ml. Lane 1: C1-inh, 125I-labeled C1s, and Patient 1 autoantibody. Lane 2: peptide 1 (2.6 µM) was preincubated with Patient 1 autoantibody at 37°C for 2 hr and then with C1-inh for a further 2 hr before addition of 125I-labeled C1s. Incubation was then continued for a further 3 hr. Lane 3: as in lane 2, except that peptide 2 (2.6 µM) was used instead of peptide 1. Lane 4: the same as in lane 2, except peptide 3 (2.6 µM) was used instead of peptide 1. Lane 5: C1-inh and 125I-labeled C1s; lane 6: I25I-labeled C1s alone. The same results were obtained with the autoantibody from Patient 2.

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