BD HorizonTM BV421 Rat Anti-Mouse IL-4

Description

Flow cytometric analysis of IL-4 expressed in activated mouse splenocytes. Splenocytes from C57BL/6 mice were enriched for CD4+ T cells by positive selection using Purified NA/LE Rat Anti-Mouse CD4 antibody-coated plates GK1.5, Cat. No.553726;10 mug/ml for 1 hr at 4C. The CD4+ T cells were harvested and stimulated with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e 145-2C11, Cat. No. 553057;10 mug/ml and soluble Purified NA/LE Hamster Anti-Mouse CD28 37.51, Cat. No. 553294; 2 mug/ml antibody and Recombinant Mouse IL-2 Cat. No. 550069; 10 ng/ml and IL-4 Cat. No. 550067; 50 ng/ml for 2 days. The cells were expanded in IL-2 and IL-4 for 3 days and then washed and stimulated 4 hr with PMA Sigma, Cat. No. P-8139; 5 ng/ml and ionomycin Sigma, Cat. No. P-8139; 500 ng in the presence of BD GolgiPlug Protein Transport Inhibitor Containing Brefeldin A Cat. No. 555029 . The activated cells were fixed with BD Cytofix Fixation Buffer Cat. No. 554655 , permeabilized using the BD Perm/Wash Permeabilization Buffer Cat. No. 554723 and then stained either with a BD Horizon BV421 Rat IgG1, kappa Isotype Control Cat. No. 562868, Left Panel or with BD Horizon BV421 Rat Anti-Mouse IL-4 antibody Cat. No. 562915, Right Panel . Two-color dot plots showing IL-4 or Ig isotype control staining versus autofluorescence were derived from gated events with the forward and light scattering characteristics of intact lymphocytes. Flow cytometry was performed using a BD LSR II Flow Cytometry System. Flow cytometric analysis of IL-4 expressed in activated mouse splenocytes. Splenocytes from C57BL/6 mice were enriched for CD4+ T cells by positive selection using Purified NA/LE Rat Anti-Mouse CD4 antibody-coated plates GK1.5, Cat. No.553726;10 mug/ml for 1 hr at 4C. The CD4+ T cells were harvested and stimulated with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e 145-2C11, Cat. No. 553057;10 mug/ml and soluble Purified NA/LE Hamster Anti-Mouse CD28 37.51, Cat. No. 553294; 2 mug/ml antibody and Recombinant Mouse IL-2 Cat. No. 550069; 10 ng/ml and IL-4 Cat. No. 550067; 50 ng/ml for 2 days. The cells were expanded in IL-2 and IL-4 for 3 days and then washed and stimulated 4 hr with PMA Sigma, Cat. No. P-8139; 5 ng/ml and ionomycin Sigma, Cat. No. P-8139; 500 ng in the presence of BD GolgiPlug Protein Transport Inhibitor Containing Brefeldin A Cat. No. 555029 . The activated cells were fixed with BD Cytofix Fixation Buffer Cat. No. 554655 , permeabilized using the BD Perm/Wash Permeabilization Buffer Cat. No. 554723 and then stained either with a BD Horizon BV421 Rat IgG1, kappa Isotype Control Cat. No. 562868, Left Panel or with BD Horizon BV421 Rat Anti-Mouse IL-4 antibody Cat. No. 562915, Right Panel . Two-color dot plots showing IL-4 or Ig isotype control staining versus autofluorescence were derived from gated events with the forward and light scattering characteristics of intact lymphocytes. Flow cytometry was performed using a BD LSR II Flow Cytometry System. Flow cytometric analysis of IL-4 expressed in activated mouse splenocytes. Splenocytes from C57BL/6 mice were enriched for CD4+ T cells by positive selection using Purified NA/LE Rat Anti-Mouse CD4 antibody-coated plates GK1.5, Cat. No.553726;10 mug/ml for 1 hr at 4C. The CD4+ T cells were harvested and stimulated with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e 145-2C11, Cat. No. 553057;10 mug/ml and soluble Purified NA/LE Hamster Anti-Mouse CD28 37.51, Cat. No. 553294; 2 mug/ml antibody and Recombinant Mouse IL-2 Cat. No. 550069; 10 ng/ml and IL-4 Cat. No. 550067; 50 ng/ml for 2 days. The cells were expanded in IL-2 and IL-4 for 3 days and then washed and stimulated 4 hr with PMA Sigma, Cat. No. P-8139; 5 ng/ml and ionomycin Sigma, Cat. No. P-8139; 500 ng in the presence of BD GolgiPlug Protein Transport Inhibitor Containing Brefeldin A Cat. No. 555029 . The activated cells were fixed with BD Cytofix Fixation Buffer Cat. No. 554655 , permeabilized using the BD Perm/Wash Permeabilization Buffer Cat. No. 554723 and then stained either with a BD Horizon BV421 Rat IgG1, kappa Isotype Control Cat. No. 562868, Left Panel or with BD Horizon BV421 Rat Anti-Mouse IL-4 antibody Cat. No. 562915, Right Panel . Two-color dot plots showing IL-4 or Ig isotype control staining versus autofluorescence were derived from gated events with the forward and light scattering characteristics of intact lymphocytes. Flow cytometry was performed using a BD LSR II Flow Cytometry System. Show More Flow cytometric analysis of IL-4 expressed in activated mouse splenocytes. Splenocytes from C57BL/6 mice were enriched for CD4+ T cells by positive selection using Purified NA/LE Rat Anti-Mouse CD4 antibody-coated plates GK1.5, Cat. No.553726;10 mug/ml for 1 hr at 4C. The CD4+ T cells were harvested and stimulated with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e 145-2C11, Cat. No. 553057;10 mug/ml and soluble Purified NA/LE Hamster Anti-Mouse CD28 37.51, Cat. No. 553294; 2 mug/ml antibody and Recombinant Mouse IL-2 Cat. No. 550069; 10 ng/ml and IL-4 Cat. No. 550067; 50 ng/ml for 2 days. The cells were expanded in IL-2 and IL-4 for 3 days and then washed and stimulated 4 hr with PMA Sigma, Cat. No. P-8139; 5 ng/ml and ionomycin Sigma, Cat. No. P-8139; 500 ng in the presence of BD GolgiPlug Protein Transport Inhibitor Containing Brefeldin A Cat. No. 555029 . The activated cells were fixed with BD Cytofix Fixation Buffer Cat. No. 554655 , permeabilized using the BD Perm/Wash Permeabilization Buffer Cat. No. 554723 and then stained either with a BD Horizon BV421 Rat IgG1, kappa Isotype Control Cat. No. 562868, Left Panel or with BD Horizon BV421 Rat Anti-Mouse IL-4 antibody Cat. No. 562915, Right Panel . Two-color dot plots showing IL-4 or Ig isotype control staining versus autofluorescence were derived from gated events with the forward and light scattering characteristics of intact lymphocytes. Flow cytometry was performed using a BD LSR II Flow Cytometry System. Show More Flow cytometric analysis of IL-4 expressed in activated mouse splenocytes. Splenocytes from C57BL/6 mice were enriched for CD4+ T cells by positive selection using Purified NA/LE Rat Anti-Mouse CD4 antibody-coated plates GK1.5, Cat. No.553726;10 mug/ml for 1 hr at 4C. The CD4+ T cells were harvested and stimulated with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e 145-2C11, Cat. No. 553057;10 mug/ml and soluble Purified NA/LE Hamster Anti-Mouse CD28 37.51, Cat. No. 553294; 2 mug/ml antibody and Recombinant Mouse IL-2 Cat. No. 550069; 10 ng/ml and IL-4 Cat. No. 550067; 50 ng/ml for 2 days. The cells were expanded in IL-2 and IL-4 for 3 days and then washed and stimulated 4 hr with PMA Sigma, Cat. No. P-8139; 5 ng/ml and ionomycin Sigma, Cat. No. P-8139; 500 ng in the presence of BD GolgiPlug Protein Transport Inhibitor Containing Brefeldin A Cat. No. 555029 . The activated cells were fixed with BD Cytofix Fixation Buffer Cat. No. 554655 , permeabilized using the BD Perm/Wash Permeabilization Buffer Cat. No. 554723 and then stained either with a BD Horizon BV421 Rat IgG1, kappa Isotype Control Cat. No. 562868, Left Panel or with BD Horizon BV421 Rat Anti-Mouse IL-4 antibody Cat. No. 562915, Right Panel . Two-color dot plots showing IL-4 or Ig isotype control staining versus autofluorescence were derived from gated events with the forward and light scattering characteristics of intact lymphocytes. Flow cytometry was performed using a BD LSR II Flow Cytometry System. Flow cytometric analysis of IL-4 expressed in activated mouse splenocytes. Splenocytes from C57BL/6 mice were enriched for CD4+ T cells by positive selection using Purified NA/LE Rat Anti-Mouse CD4 antibody-coated plates GK1.5, Cat. No.553726;10 mug/ml for 1 hr at 4C. The CD4+ T cells were harvested and stimulated with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e 145-2C11, Cat. No. 553057;10 mug/ml and soluble Purified NA/LE Hamster Anti-Mouse CD28 37.51, Cat. No. 553294; 2 mug/ml antibody and Recombinant Mouse IL-2 Cat. No. 550069; 10 ng/ml and IL-4 Cat. No. 550067; 50 ng/ml for 2 days. The cells were expanded in IL-2 and IL-4 for 3 days and then washed and stimulated 4 hr with PMA Sigma, Cat. No. P-8139; 5 ng/ml and ionomycin Sigma, Cat. No. P-8139; 500 ng in the presence of BD GolgiPlug Protein Transport Inhibitor Containing Brefeldin A Cat. No. 555029 . The activated cells were fixed with BD Cytofix Fixation Buffer Cat. No. 554655 , permeabilized using the BD Perm/Wash Permeabilization Buffer Cat. No. 554723 and then stained either with a BD Horizon BV421 Rat IgG1, kappa Isotype Control Cat. No. 562868, Left Panel or with BD Horizon BV421 Rat Anti-Mouse IL-4 antibody Cat. No. 562915, Right Panel . Two-color dot plots showing IL-4 or Ig isotype control staining versus autofluorescence were derived from gated events with the forward and light scattering characteristics of intact lymphocytes. Flow cytometry was performed using a BD LSR II Flow Cytometry System.