BD PharmingenTM PE Rat Anti-Human IL-5

Description

Detection of IL-5 expression by stimulated human CD4+ T cells. Isolated human CD4+ cells were stimulated with immobilized anti-human CD3 1 mug/ml final concentration; UCHT1, Cat. No. 555329 , soluble anti-human CD28.2 antibody 20 ng/ml final concentration; Cat. No. 555725 , recombinant human IL-2 10 ng/ml final concentration; Cat. No. 555605 and recombinant human IL-4 10 ng/ml final concentration; Cat. No. 554603 for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and recombinant human IL-4 for 3 days. The cells were then harvested and restimulated for 6 hours with PMA Sigma, Cat. #P-8139 and calcium ionophore A23187 Sigma, Cat. #C-9275 in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 . Finally, the cells were harvested, stained with 0.25 mug of PE-Cy5-anti-CD4 PE-Cy5-RPA-T4, Cat. No. 555348 , fixed, permeabilized, and subsequently stained with 20 mul PE-conjugated rat anti-human IL-5 PE-JES1-39D10, Cat. No. 559332 by following the usage section above and Pharmingens staining protocol left panel . To demonstrate specificity of staining, the binding of PE-JES1-39D10 antibody was blocked by the preincubation of the conjugated antibody with recombinant human IL-5 0.1 mug; Cat. No. 554606; middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled JES1-39D10 antibody 5 mug/Cat. No. 554487; right panel prior to staining with the PE-JEs1-39D10 antibody. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control and verified with the recombinant cytokine blocking and unlabeled blocking specificity controls. Detection of IL-5 expression by stimulated human CD4+ T cells. Isolated human CD4+ cells were stimulated with immobilized anti-human CD3 1 mug/ml final concentration; UCHT1, Cat. No. 555329 , soluble anti-human CD28.2 antibody 20 ng/ml final concentration; Cat. No. 555725 , recombinant human IL-2 10 ng/ml final concentration; Cat. No. 555605 and recombinant human IL-4 10 ng/ml final concentration; Cat. No. 554603 for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and recombinant human IL-4 for 3 days. The cells were then harvested and restimulated for 6 hours with PMA Sigma, Cat. #P-8139 and calcium ionophore A23187 Sigma, Cat. #C-9275 in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 . Finally, the cells were harvested, stained with 0.25 mug of PE-Cy5-anti-CD4 PE-Cy5-RPA-T4, Cat. No. 555348 , fixed, permeabilized, and subsequently stained with 20 mul PE-conjugated rat anti-human IL-5 PE-JES1-39D10, Cat. No. 559332 by following the usage section above and Pharmingens staining protocol left panel . To demonstrate specificity of staining, the binding of PE-JES1-39D10 antibody was blocked by the preincubation of the conjugated antibody with recombinant human IL-5 0.1 mug; Cat. No. 554606; middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled JES1-39D10 antibody 5 mug/Cat. No. 554487; right panel prior to staining with the PE-JEs1-39D10 antibody. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control and verified with the recombinant cytokine blocking and unlabeled blocking specificity controls. Detection of IL-5 expression by stimulated human CD4+ T cells. Isolated human CD4+ cells were stimulated with immobilized anti-human CD3 1 mug/ml final concentration; UCHT1, Cat. No. 555329 , soluble anti-human CD28.2 antibody 20 ng/ml final concentration; Cat. No. 555725 , recombinant human IL-2 10 ng/ml final concentration; Cat. No. 555605 and recombinant human IL-4 10 ng/ml final concentration; Cat. No. 554603 for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and recombinant human IL-4 for 3 days. The cells were then harvested and restimulated for 6 hours with PMA Sigma, Cat. #P-8139 and calcium ionophore A23187 Sigma, Cat. #C-9275 in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 . Finally, the cells were harvested, stained with 0.25 mug of PE-Cy5-anti-CD4 PE-Cy5-RPA-T4, Cat. No. 555348 , fixed, permeabilized, and subsequently stained with 20 mul PE-conjugated rat anti-human IL-5 PE-JES1-39D10, Cat. No. 559332 by following the usage section above and Pharmingens staining protocol left panel . To demonstrate specificity of staining, the binding of PE-JES1-39D10 antibody was blocked by the preincubation of the conjugated antibody with recombinant human IL-5 0.1 mug; Cat. No. 554606; middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled JES1-39D10 antibody 5 mug/Cat. No. 554487; right panel prior to staining with the PE-JEs1-39D10 antibody. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control and verified with the recombinant cytokine blocking and unlabeled blocking specificity controls. Show More Detection of IL-5 expression by stimulated human CD4+ T cells. Isolated human CD4+ cells were stimulated with immobilized anti-human CD3 1 mug/ml final concentration; UCHT1, Cat. No. 555329 , soluble anti-human CD28.2 antibody 20 ng/ml final concentration; Cat. No. 555725 , recombinant human IL-2 10 ng/ml final concentration; Cat. No. 555605 and recombinant human IL-4 10 ng/ml final concentration; Cat. No. 554603 for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and recombinant human IL-4 for 3 days. The cells were then harvested and restimulated for 6 hours with PMA Sigma, Cat. #P-8139 and calcium ionophore A23187 Sigma, Cat. #C-9275 in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 . Finally, the cells were harvested, stained with 0.25 mug of PE-Cy5-anti-CD4 PE-Cy5-RPA-T4, Cat. No. 555348 , fixed, permeabilized, and subsequently stained with 20 mul PE-conjugated rat anti-human IL-5 PE-JES1-39D10, Cat. No. 559332 by following the usage section above and Pharmingens staining protocol left panel . To demonstrate specificity of staining, the binding of PE-JES1-39D10 antibody was blocked by the preincubation of the conjugated antibody with recombinant human IL-5 0.1 mug; Cat. No. 554606; middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled JES1-39D10 antibody 5 mug/Cat. No. 554487; right panel prior to staining with the PE-JEs1-39D10 antibody. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control and verified with the recombinant cytokine blocking and unlabeled blocking specificity controls. Show More Detection of IL-5 expression by stimulated human CD4+ T cells. Isolated human CD4+ cells were stimulated with immobilized anti-human CD3 1 mug/ml final concentration; UCHT1, Cat. No. 555329 , soluble anti-human CD28.2 antibody 20 ng/ml final concentration; Cat. No. 555725 , recombinant human IL-2 10 ng/ml final concentration; Cat. No. 555605 and recombinant human IL-4 10 ng/ml final concentration; Cat. No. 554603 for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and recombinant human IL-4 for 3 days. The cells were then harvested and restimulated for 6 hours with PMA Sigma, Cat. #P-8139 and calcium ionophore A23187 Sigma, Cat. #C-9275 in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 . Finally, the cells were harvested, stained with 0.25 mug of PE-Cy5-anti-CD4 PE-Cy5-RPA-T4, Cat. No. 555348 , fixed, permeabilized, and subsequently stained with 20 mul PE-conjugated rat anti-human IL-5 PE-JES1-39D10, Cat. No. 559332 by following the usage section above and Pharmingens staining protocol left panel . To demonstrate specificity of staining, the binding of PE-JES1-39D10 antibody was blocked by the preincubation of the conjugated antibody with recombinant human IL-5 0.1 mug; Cat. No. 554606; middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled JES1-39D10 antibody 5 mug/Cat. No. 554487; right panel prior to staining with the PE-JEs1-39D10 antibody. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control and verified with the recombinant cytokine blocking and unlabeled blocking specificity controls. Detection of IL-5 expression by stimulated human CD4+ T cells. Isolated human CD4+ cells were stimulated with immobilized anti-human CD3 1 mug/ml final concentration; UCHT1, Cat. No. 555329 , soluble anti-human CD28.2 antibody 20 ng/ml final concentration; Cat. No. 555725 , recombinant human IL-2 10 ng/ml final concentration; Cat. No. 555605 and recombinant human IL-4 10 ng/ml final concentration; Cat. No. 554603 for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and recombinant human IL-4 for 3 days. The cells were then harvested and restimulated for 6 hours with PMA Sigma, Cat. #P-8139 and calcium ionophore A23187 Sigma, Cat. #C-9275 in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 . Finally, the cells were harvested, stained with 0.25 mug of PE-Cy5-anti-CD4 PE-Cy5-RPA-T4, Cat. No. 555348 , fixed, permeabilized, and subsequently stained with 20 mul PE-conjugated rat anti-human IL-5 PE-JES1-39D10, Cat. No. 559332 by following the usage section above and Pharmingens staining protocol left panel . To demonstrate specificity of staining, the binding of PE-JES1-39D10 antibody was blocked by the preincubation of the conjugated antibody with recombinant human IL-5 0.1 mug; Cat. No. 554606; middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled JES1-39D10 antibody 5 mug/Cat. No. 554487; right panel prior to staining with the PE-JEs1-39D10 antibody. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control and verified with the recombinant cytokine blocking and unlabeled blocking specificity controls.