polyclonal goat anti human il 13 antibody  (R&D Systems)

Journal: PLoS ONE

Article Title: Cytokine Induction of VCAM-1 but Not IL13Rα2 on Glioma Cells: A Tale of Two Antibodies

doi: 10.1371/journal.pone.0095123

Figure Lengend Snippet: Flow cytometric analysis of monocytic line THP-1 and various glioma lines with (A) B-D13-PE (Cell Sciences) or (B) AF146 (R&D Systems) reagents for media alone (black histogram) and cytokine (TNF/IL-4 or TNF/IL-13 overnight; red histogram) conditions. Isotype (iso-PE) and mouse anti-goat-FITC controls are shown as grey histograms. Percent positive and relative fluorescent index (RFI) of MFI cytokine/MFI media is reported for each histogram. (C) Flow cytometric detection of IL13Rα2 for D283 cells engineered to express IL13Rα2 (D283-IL13Rα2; blue histogram) and D283 parental (black histogram) stained with AF146 or B-D13-PE antibodies. All data are representative of more than three experiments each.

Article Snippet: Cell surface expression was evaluated by staining with phycoerythrin (PE)-conjugated anti-VCAM-1 (eBioscience, 12-1069-41), goat polyclonal anti-IL13Rα2 AF146 (R&D Systems) followed by detection with PE-conjugated donkey anti-goat (Novus Biologicals), PE-conjugated mouse monoclonal anti-IL13Rα2 B-D13 (Cell Sciences; lots 1107403, 1107404 and 1107405), or unconjugated B-D13 antibodies (Cell Sciences, Abcam and Santa Cruz) followed by detection with PE-conjugated goat anti-mouse (Jackson ImmunoResearch).

Techniques: Staining

Journal: PLoS ONE

Article Title: Cytokine Induction of VCAM-1 but Not IL13Rα2 on Glioma Cells: A Tale of Two Antibodies

doi: 10.1371/journal.pone.0095123

Figure Lengend Snippet: (A) D283 cells engineered to express IL13Rα2 (D283- IL13Rα2) (blue histograms) and D283 parental (black histograms) cells were evaluated by flow cytometry for expression of constitutive IL13Rα2 using the IL13Rα2-specific antibody AF146, and biotinylated recombinant human IL-13 (IL13-bio) followed by PE-conjugated strepavidin (SA-PE). Data are representative of 2 separate experiments. (B) U251T (grown in the absence of cytokines) were evaluated by flow cytometry for constitutive IL13Rα2 expression using AF146, and for binding to IL13-bio/SA-PE in the presence and absence of 10-fold molar excess of recombinant human IL-4 or IL-13. Black histograms represent staining with istoype control antibody or SA-PE alone. Data are representative of 2 separate experiments. (C) THP-1 and PBT008 grown in media alone (black histograms) or induced overnight with TNF and IL-4 (red histograms) were analyzed by flow cytometry for expression of constitutive IL13Rα2 (AF146), for expression of the induced antigen (B-D13-PE), and for binding to IL13-bio/SA-PE. Data are representative of 3 separate experiments. (D) IL13-zetakine + CD8 + CTL recognize and kill U251T glioma targets expressing constitutive IL13Rα2 (AF146-positive), but not cytokine-induced PBT003 cells (B-D13-positive). Percentage specific lysis (mean ± S.D.) of triplicate wells is depicted. *, p≤0.0002 using an unpaired Student's t-test to compare U251T vs. PBT003-4 targets. #, p>0.05 using an unpaired Student's t-test to compare PBT003-4 targets with and without overnight cytokine stimulation. Data are representative of at least 2 separate experiments.

Article Snippet: Cell surface expression was evaluated by staining with phycoerythrin (PE)-conjugated anti-VCAM-1 (eBioscience, 12-1069-41), goat polyclonal anti-IL13Rα2 AF146 (R&D Systems) followed by detection with PE-conjugated donkey anti-goat (Novus Biologicals), PE-conjugated mouse monoclonal anti-IL13Rα2 B-D13 (Cell Sciences; lots 1107403, 1107404 and 1107405), or unconjugated B-D13 antibodies (Cell Sciences, Abcam and Santa Cruz) followed by detection with PE-conjugated goat anti-mouse (Jackson ImmunoResearch).

Techniques: Flow Cytometry, Expressing, Recombinant, Binding Assay, Staining, Control, Lysis

Journal: PLoS ONE

Article Title: Cytokine Induction of VCAM-1 but Not IL13Rα2 on Glioma Cells: A Tale of Two Antibodies

doi: 10.1371/journal.pone.0095123

Figure Lengend Snippet: (A) PBT008 cells that had been cultured overnight in media alone (black histogram) versus cytokine (TNF; red histogram) conditions, (B) Parental 293T cells or 293T cells engineered to express either VCAM-1 or IL13Rα2, and (C) U251T cells were stained with VCAM-1-PE, AF146 or various B-D13 reagents – two lots of PE-conjugated B-D13 antibody (B-D13-PE; Cell Sciences) and two to three unconjugated B-D13 antibodies (B-D13-unc) purchased from either Cell Sciences (Cell Sci), Abcam or Santa Cruz as indicated. (B, C) Black histograms represent staining with istoype control antibody or SA-PE alone.

Article Snippet: Cell surface expression was evaluated by staining with phycoerythrin (PE)-conjugated anti-VCAM-1 (eBioscience, 12-1069-41), goat polyclonal anti-IL13Rα2 AF146 (R&D Systems) followed by detection with PE-conjugated donkey anti-goat (Novus Biologicals), PE-conjugated mouse monoclonal anti-IL13Rα2 B-D13 (Cell Sciences; lots 1107403, 1107404 and 1107405), or unconjugated B-D13 antibodies (Cell Sciences, Abcam and Santa Cruz) followed by detection with PE-conjugated goat anti-mouse (Jackson ImmunoResearch).

Techniques: Cell Culture, Staining, Control

Journal: PLoS ONE

Article Title: Cytokine Induction of VCAM-1 but Not IL13Rα2 on Glioma Cells: A Tale of Two Antibodies

doi: 10.1371/journal.pone.0095123

Figure Lengend Snippet: Flow cytometric analysis of monocytic line THP-1 and various glioma lines with (A) B-D13-PE (Cell Sciences) or (B) AF146 (R&D Systems) reagents for media alone (black histogram) and cytokine (TNF/IL-4 or TNF/IL-13 overnight; red histogram) conditions. Isotype (iso-PE) and mouse anti-goat-FITC controls are shown as grey histograms. Percent positive and relative fluorescent index (RFI) of MFI cytokine/MFI media is reported for each histogram. (C) Flow cytometric detection of IL13Rα2 for D283 cells engineered to express IL13Rα2 (D283-IL13Rα2; blue histogram) and D283 parental (black histogram) stained with AF146 or B-D13-PE antibodies. All data are representative of more than three experiments each.

Article Snippet: We have previously demonstrated that the goat polyclonal anti-IL13Rα2 antibody AF146 (R&D Systems) specifically recognizes human IL13Rα2 as shown by FACs, Western and qPCR .

Techniques: Staining

Journal: PLoS ONE

Article Title: Cytokine Induction of VCAM-1 but Not IL13Rα2 on Glioma Cells: A Tale of Two Antibodies

doi: 10.1371/journal.pone.0095123

Figure Lengend Snippet: (A) D283 cells engineered to express IL13Rα2 (D283- IL13Rα2) (blue histograms) and D283 parental (black histograms) cells were evaluated by flow cytometry for expression of constitutive IL13Rα2 using the IL13Rα2-specific antibody AF146, and biotinylated recombinant human IL-13 (IL13-bio) followed by PE-conjugated strepavidin (SA-PE). Data are representative of 2 separate experiments. (B) U251T (grown in the absence of cytokines) were evaluated by flow cytometry for constitutive IL13Rα2 expression using AF146, and for binding to IL13-bio/SA-PE in the presence and absence of 10-fold molar excess of recombinant human IL-4 or IL-13. Black histograms represent staining with istoype control antibody or SA-PE alone. Data are representative of 2 separate experiments. (C) THP-1 and PBT008 grown in media alone (black histograms) or induced overnight with TNF and IL-4 (red histograms) were analyzed by flow cytometry for expression of constitutive IL13Rα2 (AF146), for expression of the induced antigen (B-D13-PE), and for binding to IL13-bio/SA-PE. Data are representative of 3 separate experiments. (D) IL13-zetakine + CD8 + CTL recognize and kill U251T glioma targets expressing constitutive IL13Rα2 (AF146-positive), but not cytokine-induced PBT003 cells (B-D13-positive). Percentage specific lysis (mean ± S.D.) of triplicate wells is depicted. *, p≤0.0002 using an unpaired Student's t-test to compare U251T vs. PBT003-4 targets. #, p>0.05 using an unpaired Student's t-test to compare PBT003-4 targets with and without overnight cytokine stimulation. Data are representative of at least 2 separate experiments.

Article Snippet: We have previously demonstrated that the goat polyclonal anti-IL13Rα2 antibody AF146 (R&D Systems) specifically recognizes human IL13Rα2 as shown by FACs, Western and qPCR .

Techniques: Flow Cytometry, Expressing, Recombinant, Binding Assay, Staining, Control, Lysis

Journal: PLoS ONE

Article Title: Cytokine Induction of VCAM-1 but Not IL13Rα2 on Glioma Cells: A Tale of Two Antibodies

doi: 10.1371/journal.pone.0095123

Figure Lengend Snippet: (A) PBT008 cells that had been cultured overnight in media alone (black histogram) versus cytokine (TNF; red histogram) conditions, (B) Parental 293T cells or 293T cells engineered to express either VCAM-1 or IL13Rα2, and (C) U251T cells were stained with VCAM-1-PE, AF146 or various B-D13 reagents – two lots of PE-conjugated B-D13 antibody (B-D13-PE; Cell Sciences) and two to three unconjugated B-D13 antibodies (B-D13-unc) purchased from either Cell Sciences (Cell Sci), Abcam or Santa Cruz as indicated. (B, C) Black histograms represent staining with istoype control antibody or SA-PE alone.

Article Snippet: We have previously demonstrated that the goat polyclonal anti-IL13Rα2 antibody AF146 (R&D Systems) specifically recognizes human IL13Rα2 as shown by FACs, Western and qPCR .

Techniques: Cell Culture, Staining, Control