Journal:

Article Title: The Cysteine-Rich Region of the Entamoeba histolytica Adherence Lectin (170-Kilodalton Subunit) Is Sufficient for High-Affinity Gal/GalNAc-Specific Binding In Vitro

doi:

Figure Lengend Snippet: Immunoreactivity of 3F4 with the native lectin heavy subunit. (a) Indirect immunofluorescence of HM1:IMSS trophozoites by flow cytometric analysis. Primary antilectin MAbs 1G7, 3F4, and 8A3 (CLB was the isotype control used to measure background) were monitored by secondary goat anti-mouse-fluorescein isothiocyanate. Data depict fluorescence intensity on a logarithmic scale for 104 trophozites from a representative experiment. (b) Immunoprecipitation of biotinylated native lectin heavy subunit (170 kDa) from solubilized amebic membranes, using antilectin MAb 3F4 (IgG1) and irrelevant isotype control CLB (IgG1). Immunoprecipitates were subject to SDS-PAGE on a 7.5% gel, transferred to nitrocellulose, and probed with avidin-horseradish peroxidase.

Article Snippet: Cells were washed three times with PBS and incubated with a 1:100 dilution of goat anti-mouse-fluorescein isothiocyanate in 0.5% BSA–PBS (Bio-Rad) for 30 min on ice.

Techniques: Immunofluorescence, Fluorescence, Immunoprecipitation, SDS Page, Avidin-Biotin Assay

Journal: Journal of Biological Chemistry

Article Title: High Resolution Mapping of the Binding Site on Human IgG1 for FcγRI, FcγRII, FcγRIII, and FcRn and Design of IgG1 Variants with Improved Binding to the FcγR

doi: 10.1074/jbc.m009483200

Figure Lengend Snippet: FIG. 1. Binding of anti-IgE E27 IgG1 to human FcgR. A, binding of E27 monomers (solid circles), dimeric (solid squares), and hexameric (open squares) complexes to FcgRIIA-Arg131. B, binding of E27 mono- mers (solid circles), dimeric (solid squares), and hexameric (open squares) complexes to FcgRIIIA-Phe158. Dimers were formed by mixing E27 IgG1 and a F(ab9)2 fragment of goat anti-human k light chain at 1:0.5 molar ratio at 25 °C for 1 h (50). Hexameric complexes (i.e. trim- eric in E27 IgG1) were formed by mixing E27 IgG1 with human IgE in a 1:1 molar ratio at 25 °C for 1 h (36).

Article Snippet: Plates were blocked with assay buffer at 25 °C for 1 h. Serial 3-fold dilutions of monomeric E27 and variants (10.0–0.0045 mg/ml) were added to plates and incubated for 2 h. After washing plates with assay buffer, IgG bound to FcgRI was detected with peroxidase-conjugated F(ab9)2 fragment of goat anti-human F(ab9)2-specific IgG (Jackson ImmunoResearch) or with peroxidase-conjugated protein G (Bio-Rad).

Techniques: Binding Assay