BD PharmingenTM PE Rat Anti-Human IL-13

Description

Expression of IL-13 by stimulated human CD4+ cells. Isolated human CD4+ cells were stimulated with immobilized anti-human CD3 antibody UCHT1, 10 mug/ml for plate coating, Cat. No. 555329 , soluble anti-human CD28 antibody 20 ng/ml, Cat. No. 555725 , recombinant human IL-2 10 ng/ml, Cat. No. 554603 and recombinant human IL-4 10 ng/ml, Cat. No. 554605 for 2 days. The cells were washed and subsequently cultured in medium containing IL-2 and IL-4 for 3 days. Finally, the cells were harvested and re-stimulated for 6 hr with PMA Sigma, Cat. #P-8139; 50 ng/ml , calcium ionophore A23187 Sigma, Cat. #C-9275; 250 ng/ml in the presence of BD GolgiStop 2 muM final concentration, Cat. No. 554724 . The cells were stained with PE-Cy5-anti-CD4 PE-CY5-RPA-T4, Cat. No. 555348 fixed, permeabilized, and subsequently stained with 20 mul of PE-rat anti-human IL-13 antibody PE-JES10-5A2; Cat. No. 559328 by following the Usage section below see Right Figure, left panel . To demonstrate specificity of staining, the binding of PE-JES10-5A2 antibody was blocked by the preincubation of the conjugated antibody with excess recombinant human IL-13 see middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled JES10-5A2 antibody 5 mug, Cat. No. 554570; see right panel prior to staining with the PE-JES10-5A2 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 antibody blocking specificity controls. Expression of IL-13 by stimulated human CD4+ cells. Isolated human CD4+ cells were stimulated with immobilized anti-human CD3 antibody UCHT1, 10 mug/ml for plate coating, Cat. No. 555329 , soluble anti-human CD28 antibody 20 ng/ml, Cat. No. 555725 , recombinant human IL-2 10 ng/ml, Cat. No. 554603 and recombinant human IL-4 10 ng/ml, Cat. No. 554605 for 2 days. The cells were washed and subsequently cultured in medium containing IL-2 and IL-4 for 3 days. Finally, the cells were harvested and re-stimulated for 6 hr with PMA Sigma, Cat. #P-8139; 50 ng/ml , calcium ionophore A23187 Sigma, Cat. #C-9275; 250 ng/ml in the presence of BD GolgiStop 2 muM final concentration, Cat. No. 554724 . The cells were stained with PE-Cy5-anti-CD4 PE-CY5-RPA-T4, Cat. No. 555348 fixed, permeabilized, and subsequently stained with 20 mul of PE-rat anti-human IL-13 antibody PE-JES10-5A2; Cat. No. 559328 by following the Usage section below see Right Figure, left panel . To demonstrate specificity of staining, the binding of PE-JES10-5A2 antibody was blocked by the preincubation of the conjugated antibody with excess recombinant human IL-13 see middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled JES10-5A2 antibody 5 mug, Cat. No. 554570; see right panel prior to staining with the PE-JES10-5A2 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 antibody blocking specificity controls. Expression of IL-13 by stimulated human CD4+ cells. Isolated human CD4+ cells were stimulated with immobilized anti-human CD3 antibody UCHT1, 10 mug/ml for plate coating, Cat. No. 555329 , soluble anti-human CD28 antibody 20 ng/ml, Cat. No. 555725 , recombinant human IL-2 10 ng/ml, Cat. No. 554603 and recombinant human IL-4 10 ng/ml, Cat. No. 554605 for 2 days. The cells were washed and subsequently cultured in medium containing IL-2 and IL-4 for 3 days. Finally, the cells were harvested and re-stimulated for 6 hr with PMA Sigma, Cat. #P-8139; 50 ng/ml , calcium ionophore A23187 Sigma, Cat. #C-9275; 250 ng/ml in the presence of BD GolgiStop 2 muM final concentration, Cat. No. 554724 . The cells were stained with PE-Cy5-anti-CD4 PE-CY5-RPA-T4, Cat. No. 555348 fixed, permeabilized, and subsequently stained with 20 mul of PE-rat anti-human IL-13 antibody PE-JES10-5A2; Cat. No. 559328 by following the Usage section below see Right Figure, left panel . To demonstrate specificity of staining, the binding of PE-JES10-5A2 antibody was blocked by the preincubation of the conjugated antibody with excess recombinant human IL-13 see middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled JES10-5A2 antibody 5 mug, Cat. No. 554570; see right panel prior to staining with the PE-JES10-5A2 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 antibody blocking specificity controls. Show More Expression of IL-13 by stimulated human CD4+ cells. Isolated human CD4+ cells were stimulated with immobilized anti-human CD3 antibody UCHT1, 10 mug/ml for plate coating, Cat. No. 555329 , soluble anti-human CD28 antibody 20 ng/ml, Cat. No. 555725 , recombinant human IL-2 10 ng/ml, Cat. No. 554603 and recombinant human IL-4 10 ng/ml, Cat. No. 554605 for 2 days. The cells were washed and subsequently cultured in medium containing IL-2 and IL-4 for 3 days. Finally, the cells were harvested and re-stimulated for 6 hr with PMA Sigma, Cat. #P-8139; 50 ng/ml , calcium ionophore A23187 Sigma, Cat. #C-9275; 250 ng/ml in the presence of BD GolgiStop 2 muM final concentration, Cat. No. 554724 . The cells were stained with PE-Cy5-anti-CD4 PE-CY5-RPA-T4, Cat. No. 555348 fixed, permeabilized, and subsequently stained with 20 mul of PE-rat anti-human IL-13 antibody PE-JES10-5A2; Cat. No. 559328 by following the Usage section below see Right Figure, left panel . To demonstrate specificity of staining, the binding of PE-JES10-5A2 antibody was blocked by the preincubation of the conjugated antibody with excess recombinant human IL-13 see middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled JES10-5A2 antibody 5 mug, Cat. No. 554570; see right panel prior to staining with the PE-JES10-5A2 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 antibody blocking specificity controls. Show More Expression of IL-13 by stimulated human CD4+ cells. Isolated human CD4+ cells were stimulated with immobilized anti-human CD3 antibody UCHT1, 10 mug/ml for plate coating, Cat. No. 555329 , soluble anti-human CD28 antibody 20 ng/ml, Cat. No. 555725 , recombinant human IL-2 10 ng/ml, Cat. No. 554603 and recombinant human IL-4 10 ng/ml, Cat. No. 554605 for 2 days. The cells were washed and subsequently cultured in medium containing IL-2 and IL-4 for 3 days. Finally, the cells were harvested and re-stimulated for 6 hr with PMA Sigma, Cat. #P-8139; 50 ng/ml , calcium ionophore A23187 Sigma, Cat. #C-9275; 250 ng/ml in the presence of BD GolgiStop 2 muM final concentration, Cat. No. 554724 . The cells were stained with PE-Cy5-anti-CD4 PE-CY5-RPA-T4, Cat. No. 555348 fixed, permeabilized, and subsequently stained with 20 mul of PE-rat anti-human IL-13 antibody PE-JES10-5A2; Cat. No. 559328 by following the Usage section below see Right Figure, left panel . To demonstrate specificity of staining, the binding of PE-JES10-5A2 antibody was blocked by the preincubation of the conjugated antibody with excess recombinant human IL-13 see middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled JES10-5A2 antibody 5 mug, Cat. No. 554570; see right panel prior to staining with the PE-JES10-5A2 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 antibody blocking specificity controls. Expression of IL-13 by stimulated human CD4+ cells. Isolated human CD4+ cells were stimulated with immobilized anti-human CD3 antibody UCHT1, 10 mug/ml for plate coating, Cat. No. 555329 , soluble anti-human CD28 antibody 20 ng/ml, Cat. No. 555725 , recombinant human IL-2 10 ng/ml, Cat. No. 554603 and recombinant human IL-4 10 ng/ml, Cat. No. 554605 for 2 days. The cells were washed and subsequently cultured in medium containing IL-2 and IL-4 for 3 days. Finally, the cells were harvested and re-stimulated for 6 hr with PMA Sigma, Cat. #P-8139; 50 ng/ml , calcium ionophore A23187 Sigma, Cat. #C-9275; 250 ng/ml in the presence of BD GolgiStop 2 muM final concentration, Cat. No. 554724 . The cells were stained with PE-Cy5-anti-CD4 PE-CY5-RPA-T4, Cat. No. 555348 fixed, permeabilized, and subsequently stained with 20 mul of PE-rat anti-human IL-13 antibody PE-JES10-5A2; Cat. No. 559328 by following the Usage section below see Right Figure, left panel . To demonstrate specificity of staining, the binding of PE-JES10-5A2 antibody was blocked by the preincubation of the conjugated antibody with excess recombinant human IL-13 see middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled JES10-5A2 antibody 5 mug, Cat. No. 554570; see right panel prior to staining with the PE-JES10-5A2 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 antibody blocking specificity controls.