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 GolgiStop 2 muM final concentration, Cat. No. 554724 . The cells were stained with PE-Cy5 anti-CD4 Cat No. 555348 fixed, permeabilized, and subsequently stained with 0.25 mug of PE-rat anti-human IL-13 antibody Cat No. 562039/554571 by using Pharmingens staining protocol see left panel . To demonstrate specificity of staining, the binding of PE rat anti-human IL-13 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 Purified Rat Anti-Human IL-13 antibody 5 mug, Cat No. 554570; see right panel prior to staining with the PE rat anti-human IL-13 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 GolgiStop 2 muM final concentration, Cat. No. 554724 . The cells were stained with PE-Cy5 anti-CD4 Cat No. 555348 fixed, permeabilized, and subsequently stained with 0.25 mug of PE-rat anti-human IL-13 antibody Cat No. 562039/554571 by using Pharmingens staining protocol see left panel . To demonstrate specificity of staining, the binding of PE rat anti-human IL-13 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 Purified Rat Anti-Human IL-13 antibody 5 mug, Cat No. 554570; see right panel prior to staining with the PE rat anti-human IL-13 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 GolgiStop 2 muM final concentration, Cat. No. 554724 . The cells were stained with PE-Cy5 anti-CD4 Cat No. 555348 fixed, permeabilized, and subsequently stained with 0.25 mug of PE-rat anti-human IL-13 antibody Cat No. 562039/554571 by using Pharmingens staining protocol see left panel . To demonstrate specificity of staining, the binding of PE rat anti-human IL-13 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 Purified Rat Anti-Human IL-13 antibody 5 mug, Cat No. 554570; see right panel prior to staining with the PE rat anti-human IL-13 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 GolgiStop 2 muM final concentration, Cat. No. 554724 . The cells were stained with PE-Cy5 anti-CD4 Cat No. 555348 fixed, permeabilized, and subsequently stained with 0.25 mug of PE-rat anti-human IL-13 antibody Cat No. 562039/554571 by using Pharmingens staining protocol see left panel . To demonstrate specificity of staining, the binding of PE rat anti-human IL-13 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 Purified Rat Anti-Human IL-13 antibody 5 mug, Cat No. 554570; see right panel prior to staining with the PE rat anti-human IL-13 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 GolgiStop 2 muM final concentration, Cat. No. 554724 . The cells were stained with PE-Cy5 anti-CD4 Cat No. 555348 fixed, permeabilized, and subsequently stained with 0.25 mug of PE-rat anti-human IL-13 antibody Cat No. 562039/554571 by using Pharmingens staining protocol see left panel . To demonstrate specificity of staining, the binding of PE rat anti-human IL-13 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 Purified Rat Anti-Human IL-13 antibody 5 mug, Cat No. 554570; see right panel prior to staining with the PE rat anti-human IL-13 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 GolgiStop 2 muM final concentration, Cat. No. 554724 . The cells were stained with PE-Cy5 anti-CD4 Cat No. 555348 fixed, permeabilized, and subsequently stained with 0.25 mug of PE-rat anti-human IL-13 antibody Cat No. 562039/554571 by using Pharmingens staining protocol see left panel . To demonstrate specificity of staining, the binding of PE rat anti-human IL-13 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 Purified Rat Anti-Human IL-13 antibody 5 mug, Cat No. 554570; see right panel prior to staining with the PE rat anti-human IL-13 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.