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