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Image Search Results
Journal: The Journal of Cell Biology
Article Title: Inhibition of “self” engulfment through deactivation of myosin-II at the phagocytic synapse between human cells
doi: 10.1083/jcb.200708043
Figure Lengend Snippet: Species-specific binding of soluble human SIRPα to RBCs and CD47-coated beads. (A) Fresh human and sheep RBCs binding to soluble hSIRPα (4 μM of GST conjugate), as detected by FITC-anti-GST. “Bkgd” is obtained with RBCs plus antibody. (B) Affinity of hCD47-coated beads binding to soluble hSIRPα based on flow cytometry (see Fig. S2, available at http://www.jcb.org/cgi/content/full/jcb.200708043/DC1 ). Saturation binding fit gave the indicated dissociation constant, K d . Because this is a 3D binding constant relevant to binding in a narrow membrane gap between two cells, it is equivalent to K d ≈ 1 molecule/[10 nm x (10 μm) 2 ], which is the concentration of free SIRPα that would half-saturate CD47 on a surface. The inset shows inhibition of soluble hSIRPα binding to hCD47 beads by using anti-CD47 F(ab′) 2 generated from B6H12 antibody; similar inhibition is obtained with human RBCs.
Article Snippet: The fluorescent intensities were standardized using
Techniques: Binding Assay, Flow Cytometry, Membrane, Concentration Assay, Inhibition, Generated