The Human Phospho-Immunoreceptor Antibody Array (Catalog # ARY004) detects multiple tyrosine-phosphorylated immunoreceptors in lysates prepared from cells activated by antibody-mediated cross-linking. A. THP-1 cells (human monocytic leukemia cell line) were incubated with goat anti-Fc? RIIA antibody (Catalog # AF1875) or control goat IgG (Catalog # AB-108-C) followed by incubation with a donkey anti-goat IgG antibody (Catalog # AF109) for five minutes. B. Jurkat cells were incubated with a mouse monoclonal anti-CD3e antibody (Catalog # MAB100) or mouse IgG1 isotype control (Catalog # MAB002) followed by incubation with a goat anti-mouse antibody (Catalog # AF007) for 5 minutes.
All arrays were incubated with 100 µg of lysate and phosphorylation detected by chemiluminescence. Array signals from scanned X-ray film images were analyzed using image analysis software. Changes in the phosphorylation of several immunoreceptors (table) are highlighted in the blots and histograms.