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Multicolor flow cytometric analysis of IL-9 expression by unstimulated and activated mouse spleen cells. Mouse spleen cells were either unstimulated (Left Panel) or stimulated in a tissue culture plate coated with Anti-Mouse CD3e and soluble Anti-Mouse CD28 antibodies along with Recombinant Mouse IL-2, IL-4, and TGF-β proteins and Anti-Mouse IFN-γ antibody for 4 days. On day 4 the cells were harvested and restimulated with Phorbol 12-Myristate 13-Acetate (PMA; Sigma P-8139) plus Ionomycin (Sigma; I-0634) in the presence of BD GolgiStop™ Protein Transport Inhibitor for 5 hours (Right Panel). The cells were then fixed and permeabilized using a BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit followed by staining with PE Armenian Hamster anti-Mouse IL-9 (Cat. No. 561463) and Alexa Fluor® 647 Rat Anti-Mouse CD4 (Cat. No. 557681). Two-color flow cytometric dot plots showing the correlated expression patterns of CD4 versus IL-9 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System. Other compatible fixation and permeabilization treatments are listed in the Suggested Companion Products.
Multicolor flow cytometric analysis of IL-9 expression by unstimulated and activated mouse spleen cells. Mouse spleen cells were either unstimulated (Left Panel) or stimulated in a tissue culture plate coated with Anti-Mouse CD3e and soluble Anti-Mouse CD28 antibodies along with Recombinant Mouse IL-2, IL-4, and TGF-β proteins and Anti-Mouse IFN-γ antibody for 4 days. On day 4 the cells were harvested and restimulated with Phorbol 12-Myristate 13-Acetate (PMA; Sigma P-8139) plus Ionomycin (Sigma; I-0634) in the presence of BD GolgiStop™ Protein Transport Inhibitor for 5 hours (Right Panel). The cells were then fixed and permeabilized using a BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit followed by staining with PE Armenian Hamster anti-Mouse IL-9 (Cat. No. 561463) and Alexa Fluor® 647 Rat Anti-Mouse CD4 (Cat. No. 557681). Two-color flow cytometric dot plots showing the correlated expression patterns of CD4 versus IL-9 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System. Other compatible fixation and permeabilization treatments are listed in the Suggested Companion Products.
Multicolor flow cytometric analysis of IL-9 expression by unstimulated and activated mouse spleen cells. Mouse spleen cells were either unstimulated (Left Panel) or stimulated in a tissue culture plate coated with Anti-Mouse CD3e and soluble Anti-Mouse CD28 antibodies along with Recombinant Mouse IL-2, IL-4, and TGF-β proteins and Anti-Mouse IFN-γ antibody for 4 days. On day 4 the cells were harvested and restimulated with Phorbol 12-Myristate 13-Acetate (PMA; Sigma P-8139) plus Ionomycin (Sigma; I-0634) in the presence of BD GolgiStop™ Protein Transport Inhibitor for 5 hours (Right Panel). The cells were then fixed and permeabilized using a BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit followed by staining with PE Armenian Hamster anti-Mouse IL-9 (Cat. No. 561463) and Alexa Fluor® 647 Rat Anti-Mouse CD4 (Cat. No. 557681). Two-color flow cytometric dot plots showing the correlated expression patterns of CD4 versus IL-9 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System. Other compatible fixation and permeabilization treatments are listed in the Suggested Companion Products.
Multicolor flow cytometric analysis of IL-9 expression by unstimulated and activated mouse spleen cells. Mouse spleen cells were either unstimulated (Left Panel) or stimulated in a tissue culture plate coated with Anti-Mouse CD3e and soluble Anti-Mouse CD28 antibodies along with Recombinant Mouse IL-2, IL-4, and TGF-β proteins and Anti-Mouse IFN-γ antibody for 4 days. On day 4 the cells were harvested and restimulated with Phorbol 12-Myristate 13-Acetate (PMA; Sigma P-8139) plus Ionomycin (Sigma; I-0634) in the presence of BD GolgiStop™ Protein Transport Inhibitor for 5 hours (Right Panel). The cells were then fixed and permeabilized using a BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit followed by staining with PE Armenian Hamster anti-Mouse IL-9 (Cat. No. 561463) and Alexa Fluor® 647 Rat Anti-Mouse CD4 (Cat. No. 557681). Two-color flow cytometric dot plots showing the correlated expression patterns of CD4 versus IL-9 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System. Other compatible fixation and permeabilization treatments are listed in the Suggested Companion Products.
Multicolor flow cytometric analysis of IL-9 expression by unstimulated and activated mouse spleen cells. Mouse spleen cells were either unstimulated (Left Panel) or stimulated in a tissue culture plate coated with Anti-Mouse CD3e and soluble Anti-Mouse CD28 antibodies along with Recombinant Mouse IL-2, IL-4, and TGF-β proteins and Anti-Mouse IFN-γ antibody for 4 days. On day 4 the cells were harvested and restimulated with Phorbol 12-Myristate 13-Acetate (PMA; Sigma P-8139) plus Ionomycin (Sigma; I-0634) in the presence of BD GolgiStop™ Protein Transport Inhibitor for 5 hours (Right Panel). The cells were then fixed and permeabilized using a BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit followed by staining with PE Armenian Hamster anti-Mouse IL-9 (Cat. No. 561463) and Alexa Fluor® 647 Rat Anti-Mouse CD4 (Cat. No. 557681). Two-color flow cytometric dot plots showing the correlated expression patterns of CD4 versus IL-9 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System. Other compatible fixation and permeabilization treatments are listed in the Suggested Companion Products.
Multicolor flow cytometric analysis of IL-9 expression by unstimulated and activated mouse spleen cells. Mouse spleen cells were either unstimulated (Left Panel) or stimulated in a tissue culture plate coated with Anti-Mouse CD3e and soluble Anti-Mouse CD28 antibodies along with Recombinant Mouse IL-2, IL-4, and TGF-β proteins and Anti-Mouse IFN-γ antibody for 4 days. On day 4 the cells were harvested and restimulated with Phorbol 12-Myristate 13-Acetate (PMA; Sigma P-8139) plus Ionomycin (Sigma; I-0634) in the presence of BD GolgiStop™ Protein Transport Inhibitor for 5 hours (Right Panel). The cells were then fixed and permeabilized using a BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit followed by staining with PE Armenian Hamster anti-Mouse IL-9 (Cat. No. 561463) and Alexa Fluor® 647 Rat Anti-Mouse CD4 (Cat. No. 557681). Two-color flow cytometric dot plots showing the correlated expression patterns of CD4 versus IL-9 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System. Other compatible fixation and permeabilization treatments are listed in the Suggested Companion Products.
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This fluorescent antibody is suitable for intracellular staining of mouse leukocytes using BD Cytofix/Cytoperm™ Reagents or BD Phosflow™ Fix Buffer I and Perm/Wash Buffer I (please see Suggested Companion Products).
The D9302C12 monoclonal antibody specifically binds to the multifunctional mouse cytokine, Interleukin-9 (IL-9). IL-9 is a 126 amino acid-long glycoprotein that is produced by various subsets of activated CD4+ T cells. IL-9 acts on target cells by binding to and signaling through the heterodimeric IL-9 receptor (IL-9R) complex that is comprised of transmembrane IL-9 receptor alpha (IL-9Rα) and common gamma chain (γc) subunits. IL-9 can promote the survival, growth, proliferation and/or differentiation of various cell types including thymocytes, T cells, B cells, mast cells, and hematopoietic progenitor cells. IL-9 can augment IL-4-induced IgE and IgG1 production from lipopolysaccharide-primed mouse B cells and induce granzyme and high-affinity IgE receptor gene expression by mouse T helper cell clones and mast cell lines. IL-9 plays an important role in vivo in helminth elimination. The D9302C12 antibody neutralizes mouse IL-9 bioactivity.
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