Journal: Chemical Science
Article Title: A (poly)Pro tip for preserving native disulfide connectivity during thiol–maleimide bioconjugation of disulfide-rich peptides
doi: 10.1039/d5sc08821f
Figure Lengend Snippet: (A) ELISA cell assay showing the interaction between decreasing concentrations of polyclonal IgG and HEK cells expressing LINGO-1. Absorbance values at 450 nm were background-corrected by subtracting the signal from non-transfected HEK cells. (B) Superposition of the [1–20] region for LINGO-1-to-4 homologs shows that all Cys residues are conserved; additional conserved residues are highlighted in dark grey, whereas similar residues—defined by comparable hydrophilicity ( e.g. , Ser vs. Thr) or charge ( e.g. , Lys, Arg, or His)—are shown in light grey. (C) Western blot detail in which polyclonal IgG was used for staining HEK cells and LINGO homologs 1–4. Anti-GAPDH staining is shown at the bottom as a loading control.
Article Snippet: Remarkably, our polyclonal IgG detected LINGO-1 at a five-fold higher dilution than the commercial polyclonal IgG antibody AF3086 In a previous study, screening of all commercially available IgGs identified polyclonal IgG AF3086 as the most suitable for co-immunoprecipitation studies. (R&D Systems), while also yielding stronger overall staining intensity (Fig. S52 and S53).
Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Transfection, Western Blot, Staining, Control