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. 2018 Oct 30;293(51):19645–19658. doi: 10.1074/jbc.RA118.005676

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© 2018 Huang et al.

Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc.

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Figure 4. — Specificity of rabbit polyclonal antibody toward C-terminal heptapeptide of soluble Siglec-14. A, N-terminal His₆-tagged recombinant soluble Siglec-14 proteins (full-length, C-terminally truncated (ΔC), or with R119A mutation) were subjected to nonreducing SDS-PAGE/Western blotting and stained with mouse mAb clone 40-1 (pan-Siglec-14 antibody; left panel) or with rabbit polyclonal antibody against soluble Siglec-14 C-terminal heptapeptide that is unique to the isoform generated by alternative mRNA splicing (right panel). Filled and open arrowheads indicate monomer and dimers of soluble Siglec-14, respectively. B, ELISA standard curves (capture: mouse mAb, clone 194128; detection: clone 40-1 (left panel) or rabbit polyclonal antibody against C-terminal heptapeptide (right panel)) for the standard proteins, N-terminal His₆-tagged soluble Siglec-14 (solid circle), and N-terminal His₆-tagged and C-terminal truncated soluble Siglec-14 (open circle). Error bars represent standard deviation of technical triplicates.