siglec 8 pe fab7975p  (R&D Systems)

Journal: Frontiers in Immunology

Article Title: Human Peripheral Blood Eosinophils Express High Levels of the Purinergic Receptor P2X4

doi: 10.3389/fimmu.2019.02074

Figure Lengend Snippet: Anti-hP2X4 recognizes hP2X4 in immunoprecipitation experiments, but not in western blot. (A) Left panel. Western blot using mAb19 , mAb27 , rabbit anti-rat-P2X4 polyclonal antibodies from Alomone or our mouse anti-hP2X4 polyclonal antibodies , performed on total cell lysate of HEK293 cells transfected with hP2X4. Blots were revealed using anti-mouse and anti-rabbit HRP Abs. Right panel. hP2X4 was immuno-precipitated from the same lysate of HEK293 cells expressing human P2X4, using mAb19 or mAb27 , and revealed by western blotting with rabbit anti-P2X4 polyclonal antibodies. Negative controls are shown in lanes 7 and 8: immunoprecipitation of non transfected HEK293 cell lysates with mAb19 (lane 7) and mAb27 (lane 8). Immunoprecipitation of P2X4 from cell lysate of HEK293 cells transfected with hP2X4 using rabbit anti-rat P2X4 polyclonal antibodies is shown in lane 9. (B) Immunoprecipitation assays using mAb27 (IgG2b; lanes 1, 4, 7) or mAb29 (IgM; lanes 2, 5, 8) or anti-IgM (lanes 3 and 6) from HEK cells overexpressing hP2X4-mcherry (lanes 1–3), mouse P2X4-mcherry (lanes 4–6), or non transfected (lanes 7, 8). P2X4 was revealed by western blotting with rabbit anti-P2X4 polyclonal antibodies. Arrows indicate hP2X4 or moP2X4 bands.

Article Snippet: Sections were stained with anti-hP2X4 mAb27-FITC (1:1,600; IgG2b), or mouse anti-h-Siglec-8-PE mAb (1:10; IgG1; from BioLegend clone 7C9).

Techniques: Immunoprecipitation, Western Blot, Transfection, Expressing

Journal: Frontiers in Immunology

Article Title: Human Peripheral Blood Eosinophils Express High Levels of the Purinergic Receptor P2X4

doi: 10.3389/fimmu.2019.02074

Figure Lengend Snippet: Anti-hP2X4 mAb27 recognizes mouse and rat P2X4 in immunohistochemistry assays. (A) Staining of HEK cells transfected with hP2X4 using mAb27 (IgG2b/κ). Scale bar: 15 μm. Staining of HEK cells expressing hP2X4 with secondary Ab alone did not show positive signal . (B) HEK cells expressing mouse (left panel) or human (right panel) P2X4 were stained by anti-hP2X4 mAb27-FITC (IgG 2b/κ) diluted 1:1,600 (red line) or 1:800 (blue line). The black line corresponds to staining with the isotype control. (C) Primary culture of rat hippocampus neurons with one amoebaean-like glial cell (arrow) was stained with anti-hP2X4 mAb27 (IgG2b/κ). Scale bar: 30 μm. Staining of rat hippocampus neurons with secondary antibodies alone was negative .

Article Snippet: Sections were stained with anti-hP2X4 mAb27-FITC (1:1,600; IgG2b), or mouse anti-h-Siglec-8-PE mAb (1:10; IgG1; from BioLegend clone 7C9).

Techniques: Immunohistochemistry, Staining, Transfection, Expressing, Control