BD PharmingenTM PE Rat Anti-Human IL-4

Description

Expression of IL-4 by stimulated human peripheral blood mononuclear cells PBMC . Human PBMC were stimulated with soluble anti-human CD3 antibody 1 mug/ml final concentration; Cat. No. 555329 , recombinant human IL-2 10 ng/ml final concentration; Cat. No. 554603 and recombinant human IL-4 10 ng/ml final concentration; Cat. No. 554605 for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and recombinant human IL-4 for 3 days. Finally, the cells were harvested and stimulated for 6 hours with PMA Sigma, Cat. #P-8139 and calcium ionophore A23187 Sigma in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 . The cells were harvested, stained with PE-Cy5 formerly known as Cy-Chrome -anti CD4 Cat. No. 555348 , fixed, permeabilized, and subsequently stained with 0.05 mug of PE-rat anti-human IL-4 antibody PE-MP4-25D2, Cat. No. 554485 by using PharMingens staining protocol left panel . To demonstrate specificity of staining, the binding of PE-MP4-25D2 antibody was blocked by preincubation of the fixed/permeabilized cells with unlabelled MP4-25D2 antibody 2.5 mug, Cat. No. 554482; right panel prior to staining. The quadrant markers for the bivariate dot plot were set based on the autofluorescence controls and verified using unlabelled antibody blocking controls. Expression of IL-4 by stimulated human peripheral blood mononuclear cells PBMC . Human PBMC were stimulated with soluble anti-human CD3 antibody 1 mug/ml final concentration; Cat. No. 555329 , recombinant human IL-2 10 ng/ml final concentration; Cat. No. 554603 and recombinant human IL-4 10 ng/ml final concentration; Cat. No. 554605 for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and recombinant human IL-4 for 3 days. Finally, the cells were harvested and stimulated for 6 hours with PMA Sigma, Cat. #P-8139 and calcium ionophore A23187 Sigma in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 . The cells were harvested, stained with PE-Cy5 formerly known as Cy-Chrome -anti CD4 Cat. No. 555348 , fixed, permeabilized, and subsequently stained with 0.05 mug of PE-rat anti-human IL-4 antibody PE-MP4-25D2, Cat. No. 554485 by using PharMingens staining protocol left panel . To demonstrate specificity of staining, the binding of PE-MP4-25D2 antibody was blocked by preincubation of the fixed/permeabilized cells with unlabelled MP4-25D2 antibody 2.5 mug, Cat. No. 554482; right panel prior to staining. The quadrant markers for the bivariate dot plot were set based on the autofluorescence controls and verified using unlabelled antibody blocking controls. Expression of IL-4 by stimulated human peripheral blood mononuclear cells PBMC . Human PBMC were stimulated with soluble anti-human CD3 antibody 1 mug/ml final concentration; Cat. No. 555329 , recombinant human IL-2 10 ng/ml final concentration; Cat. No. 554603 and recombinant human IL-4 10 ng/ml final concentration; Cat. No. 554605 for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and recombinant human IL-4 for 3 days. Finally, the cells were harvested and stimulated for 6 hours with PMA Sigma, Cat. #P-8139 and calcium ionophore A23187 Sigma in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 . The cells were harvested, stained with PE-Cy5 formerly known as Cy-Chrome -anti CD4 Cat. No. 555348 , fixed, permeabilized, and subsequently stained with 0.05 mug of PE-rat anti-human IL-4 antibody PE-MP4-25D2, Cat. No. 554485 by using PharMingens staining protocol left panel . To demonstrate specificity of staining, the binding of PE-MP4-25D2 antibody was blocked by preincubation of the fixed/permeabilized cells with unlabelled MP4-25D2 antibody 2.5 mug, Cat. No. 554482; right panel prior to staining. The quadrant markers for the bivariate dot plot were set based on the autofluorescence controls and verified using unlabelled antibody blocking controls. Show More Expression of IL-4 by stimulated human peripheral blood mononuclear cells PBMC . Human PBMC were stimulated with soluble anti-human CD3 antibody 1 mug/ml final concentration; Cat. No. 555329 , recombinant human IL-2 10 ng/ml final concentration; Cat. No. 554603 and recombinant human IL-4 10 ng/ml final concentration; Cat. No. 554605 for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and recombinant human IL-4 for 3 days. Finally, the cells were harvested and stimulated for 6 hours with PMA Sigma, Cat. #P-8139 and calcium ionophore A23187 Sigma in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 . The cells were harvested, stained with PE-Cy5 formerly known as Cy-Chrome -anti CD4 Cat. No. 555348 , fixed, permeabilized, and subsequently stained with 0.05 mug of PE-rat anti-human IL-4 antibody PE-MP4-25D2, Cat. No. 554485 by using PharMingens staining protocol left panel . To demonstrate specificity of staining, the binding of PE-MP4-25D2 antibody was blocked by preincubation of the fixed/permeabilized cells with unlabelled MP4-25D2 antibody 2.5 mug, Cat. No. 554482; right panel prior to staining. The quadrant markers for the bivariate dot plot were set based on the autofluorescence controls and verified using unlabelled antibody blocking controls. Show More Expression of IL-4 by stimulated human peripheral blood mononuclear cells PBMC . Human PBMC were stimulated with soluble anti-human CD3 antibody 1 mug/ml final concentration; Cat. No. 555329 , recombinant human IL-2 10 ng/ml final concentration; Cat. No. 554603 and recombinant human IL-4 10 ng/ml final concentration; Cat. No. 554605 for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and recombinant human IL-4 for 3 days. Finally, the cells were harvested and stimulated for 6 hours with PMA Sigma, Cat. #P-8139 and calcium ionophore A23187 Sigma in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 . The cells were harvested, stained with PE-Cy5 formerly known as Cy-Chrome -anti CD4 Cat. No. 555348 , fixed, permeabilized, and subsequently stained with 0.05 mug of PE-rat anti-human IL-4 antibody PE-MP4-25D2, Cat. No. 554485 by using PharMingens staining protocol left panel . To demonstrate specificity of staining, the binding of PE-MP4-25D2 antibody was blocked by preincubation of the fixed/permeabilized cells with unlabelled MP4-25D2 antibody 2.5 mug, Cat. No. 554482; right panel prior to staining. The quadrant markers for the bivariate dot plot were set based on the autofluorescence controls and verified using unlabelled antibody blocking controls. Expression of IL-4 by stimulated human peripheral blood mononuclear cells PBMC . Human PBMC were stimulated with soluble anti-human CD3 antibody 1 mug/ml final concentration; Cat. No. 555329 , recombinant human IL-2 10 ng/ml final concentration; Cat. No. 554603 and recombinant human IL-4 10 ng/ml final concentration; Cat. No. 554605 for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and recombinant human IL-4 for 3 days. Finally, the cells were harvested and stimulated for 6 hours with PMA Sigma, Cat. #P-8139 and calcium ionophore A23187 Sigma in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 . The cells were harvested, stained with PE-Cy5 formerly known as Cy-Chrome -anti CD4 Cat. No. 555348 , fixed, permeabilized, and subsequently stained with 0.05 mug of PE-rat anti-human IL-4 antibody PE-MP4-25D2, Cat. No. 554485 by using PharMingens staining protocol left panel . To demonstrate specificity of staining, the binding of PE-MP4-25D2 antibody was blocked by preincubation of the fixed/permeabilized cells with unlabelled MP4-25D2 antibody 2.5 mug, Cat. No. 554482; right panel prior to staining. The quadrant markers for the bivariate dot plot were set based on the autofluorescence controls and verified using unlabelled antibody blocking controls.