Human IL-4R alpha Antibody

Detection of IL‑4 R alpha in Daudi Human Cell Line by Flow Cytometry. Daudi human Burkitt's lymphoma cell line was stained with Mouse Anti-Human IL-4 Ra Monoclonal Antibody (Catalog # MAB230, filled histogram) or isotype control antibody (Catalog # MAB003, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B).

IL‑4 R alpha in Human Lymph Node. IL-4 Ra was detected in immersion fixed paraffin-embedded sections of human lymph node using Mouse Anti-Human IL-4 Ra Monoclonal Antibody (Catalog # MAB230) at 25 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to plasma cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

IL‑4 R alpha in HDLM‑2 Human Cell Line. IL-4 Ra was detected in immersion fixed HDLM-2 human Hodgkin's lymphoma cell line using Mouse Anti-Human IL-4 Ra Monoclonal Antibody (Catalog # MAB230) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Cell Proliferation Induced by IL‑4 and Neutralization by Human IL‑4 R alpha Antibody. Recombinant Human IL‑4 (Catalog # 204-IL) stimulates proliferation in the TF‑1 human erythroleukemic cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human IL‑4 (0.2 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human IL‑4 Ra Monoclonal Antibody (Catalog # MAB230). The ND₅₀ is typically 3-6 ng/mL.

Detection of Human IL-4R alpha by Flow Cytometry In vitro binding assays of [V14]6 and [AP1-V12]6 polymer.(A, B) H226, (C, D) MDA-MB-231 and (E, F) H460 cells were incubated with 10 µM of [V14]6, [AP1-V12]6 and AP1 for 1 h at 4°C. Cell binding was determined using flow cytometry. Histograms are representative of three independent experiments. Graphical bars (on right) represent the percent of Alexa 488 labeled polymer bound to cells as mean ±SD of data obtained from three separate experiments performed in triplicates. ***P<0.0001, **P<0.001, and *P<0.05, one-way ANOVA; n = 3. (G, H) H226 cells (3×105 cells) were pre- incubated with different concentrations (1, 5 and 10 µg/mL) of anti-IL-4 receptor antibody followed by 1 h incubation with 10 µM Alexa-labeled [AP1-V12]6 at 4°C. The cells were further suspended in 300 µL of PBS after washing and analyzed using flow cytometry. Histograms are representative of three independent experiments. Graphical bars represent the percent of Alexa 488 labeled polymer bound to cells as mean ±SD of data obtained from three separate experiments performed in triplicates. ***P<0.0001, One way ANOVA; n = 3. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0081891), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human IL-4R alpha by Immunocytochemistry/Immunofluorescence In vivo, ex vivo imaging and biodistribution of [AP1-V12]6 polymers.(A) Cy5.5-labeled [V14]6 and [AP1-V12]6 (8 mg/kg) were intravenously injected into MDA-MB-231 tumor xenografted nude mice. Biodistribution was determined by collecting in vivo fluorescence images at different time points. Scale bar indicates normalized fluorescence intensity (NC). Representative optical images of three experiments. (B) Quantitation of fluorescence intensities in tumor sites at respective time points (n = 3). (C) Analysis of fluorescence intensities for tumors and organs from ex vivo images (n = 3). (D) Histological analysis of [AP1-V12]6 polymer (red) localization in tumors. Nuclei were stained with DAPI (blue), and IL-4R expression on cells was visualized by anti-IL-4 receptor antibody staining (green). Representative confocal images of three experiments (scale bar 50 µm). Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0081891), licensed under a CC-BY license. Not internally tested by R&D Systems.