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Image Search Results
Journal: The Journal of Experimental Medicine
Article Title: Continual Low-Level Activation of the Classical Complement Pathway
doi:
Figure Lengend Snippet: Detection of C3 fragments in mouse and human blood following incubation ex vivo at 37°C. (A) The level of C3 fragments in mouse blood collected directly into heparin and EDTA (T 0 ) or collected into heparin and incubated at 37°C for 10 min before addition of EDTA (T 10 ). The histograms represent the amount of C3 fragments detected on B220 + B cells following incubation of splenic lymphocytes with T 0 plasma (filled histogram) or T 10 plasma (open histogram), as measured by immunofluorescence flow cytometry. (B) Western blot analysis of C3 fragments in mouse plasma, as collected in A. C3 was immunoprecipitated from plasma samples with polyclonal anti–mouse C3 antibody conjugated Sepharose beads, and the Western blot was probed with biotin-labeled polyclonal anti–mouse C3 Ab and developed using streptavidin-HRP. Position of intact C3α and C3β chains and the C3dg fragments, that bind to CR1/2 on B cells (A), is indicated. (C) The level of C3 fragments in human blood collected directly into heparin and EDTA (T 0 ) or into heparin and incubated at 37°C for 30 min before addition of EDTA (T 30 ). The histograms represent the amount of C3 (solid line) or C3d (broken line) fragments detected on Raji cells after incubation with T 0 plasma (filled histogram) or T 30 plasma (open histograms), as measured by immunofluorescence flow cytometry. (D) The rate of formation of C3 fragments in blood from C57BL/6 mice collected into either heparin (□) or hirudin (▪) is depicted, as measured by the binding of C3 fragments to B220 + spleen cells using immunofluorescence flow cytometry, as shown in A. Error bars represent ± SD of means of triplicate assays.
Article Snippet: For human plasma samples, Raji cells were incubated with either a
Techniques: Incubation, Ex Vivo, Clinical Proteomics, Immunofluorescence, Flow Cytometry, Western Blot, Immunoprecipitation, Labeling, Binding Assay