BD PharmingenTM FITC Rat IgG2a, kappa Isotype Control

Description

Expression of IL-2 by stimulated CD3+ human PBMC. Human PBMC were stimulated for 6 hours with PMA Sigma and calcium ionophore A23187 Sigma in the presence of GolgiStop 2 mM final concentration; Cat. No. 554714 . The PBMC were stained with PE-Cy5-anti-CD3 PE-CY5 UCHT1, Cat. No 555334 , fixed, permeabilized, and subsequently stained with 0.25 mg of FITC-Rat IgG2a, kappa anti-human IL-2 antibody FITC-MQ1-17H12, Cat. No 554565; left panel or 0.25 mg FITC-R35-95 isotype control immunoglobulin FITC-R35-95, Cat. No. 554688; right panel using Pharmingens staining protocol. To demonstrate specificity of staining, the binding of FITC-MQ1-17H12 was blocked by the preincubation of the conjugated antibody with molar excess of recombinant human IL-2 Cat. No.554603; data not shown , and by preincubation of the fixed/permeabilized cells with an excess of the unlabelled MQ1-17H12 antibody Cat. No. 554563; data not shown prior to staining with the FITC-MQ1-17H12 antibody. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the recombinant cytokine blocking and unlabelled antibody blocking specificity controls. Expression of IL-2 by stimulated CD3+ human PBMC. Human PBMC were stimulated for 6 hours with PMA Sigma and calcium ionophore A23187 Sigma in the presence of GolgiStop 2 mM final concentration; Cat. No. 554714 . The PBMC were stained with PE-Cy5-anti-CD3 PE-CY5 UCHT1, Cat. No 555334 , fixed, permeabilized, and subsequently stained with 0.25 mg of FITC-Rat IgG2a, kappa anti-human IL-2 antibody FITC-MQ1-17H12, Cat. No 554565; left panel or 0.25 mg FITC-R35-95 isotype control immunoglobulin FITC-R35-95, Cat. No. 554688; right panel using Pharmingens staining protocol. To demonstrate specificity of staining, the binding of FITC-MQ1-17H12 was blocked by the preincubation of the conjugated antibody with molar excess of recombinant human IL-2 Cat. No.554603; data not shown , and by preincubation of the fixed/permeabilized cells with an excess of the unlabelled MQ1-17H12 antibody Cat. No. 554563; data not shown prior to staining with the FITC-MQ1-17H12 antibody. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the recombinant cytokine blocking and unlabelled antibody blocking specificity controls. Expression of IL-2 by stimulated CD3+ human PBMC. Human PBMC were stimulated for 6 hours with PMA Sigma and calcium ionophore A23187 Sigma in the presence of GolgiStop 2 mM final concentration; Cat. No. 554714 . The PBMC were stained with PE-Cy5-anti-CD3 PE-CY5 UCHT1, Cat. No 555334 , fixed, permeabilized, and subsequently stained with 0.25 mg of FITC-Rat IgG2a, kappa anti-human IL-2 antibody FITC-MQ1-17H12, Cat. No 554565; left panel or 0.25 mg FITC-R35-95 isotype control immunoglobulin FITC-R35-95, Cat. No. 554688; right panel using Pharmingens staining protocol. To demonstrate specificity of staining, the binding of FITC-MQ1-17H12 was blocked by the preincubation of the conjugated antibody with molar excess of recombinant human IL-2 Cat. No.554603; data not shown , and by preincubation of the fixed/permeabilized cells with an excess of the unlabelled MQ1-17H12 antibody Cat. No. 554563; data not shown prior to staining with the FITC-MQ1-17H12 antibody. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the recombinant cytokine blocking and unlabelled antibody blocking specificity controls. Show More Expression of IL-2 by stimulated CD3+ human PBMC. Human PBMC were stimulated for 6 hours with PMA Sigma and calcium ionophore A23187 Sigma in the presence of GolgiStop 2 mM final concentration; Cat. No. 554714 . The PBMC were stained with PE-Cy5-anti-CD3 PE-CY5 UCHT1, Cat. No 555334 , fixed, permeabilized, and subsequently stained with 0.25 mg of FITC-Rat IgG2a, kappa anti-human IL-2 antibody FITC-MQ1-17H12, Cat. No 554565; left panel or 0.25 mg FITC-R35-95 isotype control immunoglobulin FITC-R35-95, Cat. No. 554688; right panel using Pharmingens staining protocol. To demonstrate specificity of staining, the binding of FITC-MQ1-17H12 was blocked by the preincubation of the conjugated antibody with molar excess of recombinant human IL-2 Cat. No.554603; data not shown , and by preincubation of the fixed/permeabilized cells with an excess of the unlabelled MQ1-17H12 antibody Cat. No. 554563; data not shown prior to staining with the FITC-MQ1-17H12 antibody. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the recombinant cytokine blocking and unlabelled antibody blocking specificity controls. Show More Expression of IL-2 by stimulated CD3+ human PBMC. Human PBMC were stimulated for 6 hours with PMA Sigma and calcium ionophore A23187 Sigma in the presence of GolgiStop 2 mM final concentration; Cat. No. 554714 . The PBMC were stained with PE-Cy5-anti-CD3 PE-CY5 UCHT1, Cat. No 555334 , fixed, permeabilized, and subsequently stained with 0.25 mg of FITC-Rat IgG2a, kappa anti-human IL-2 antibody FITC-MQ1-17H12, Cat. No 554565; left panel or 0.25 mg FITC-R35-95 isotype control immunoglobulin FITC-R35-95, Cat. No. 554688; right panel using Pharmingens staining protocol. To demonstrate specificity of staining, the binding of FITC-MQ1-17H12 was blocked by the preincubation of the conjugated antibody with molar excess of recombinant human IL-2 Cat. No.554603; data not shown , and by preincubation of the fixed/permeabilized cells with an excess of the unlabelled MQ1-17H12 antibody Cat. No. 554563; data not shown prior to staining with the FITC-MQ1-17H12 antibody. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the recombinant cytokine blocking and unlabelled antibody blocking specificity controls. Expression of IL-2 by stimulated CD3+ human PBMC. Human PBMC were stimulated for 6 hours with PMA Sigma and calcium ionophore A23187 Sigma in the presence of GolgiStop 2 mM final concentration; Cat. No. 554714 . The PBMC were stained with PE-Cy5-anti-CD3 PE-CY5 UCHT1, Cat. No 555334 , fixed, permeabilized, and subsequently stained with 0.25 mg of FITC-Rat IgG2a, kappa anti-human IL-2 antibody FITC-MQ1-17H12, Cat. No 554565; left panel or 0.25 mg FITC-R35-95 isotype control immunoglobulin FITC-R35-95, Cat. No. 554688; right panel using Pharmingens staining protocol. To demonstrate specificity of staining, the binding of FITC-MQ1-17H12 was blocked by the preincubation of the conjugated antibody with molar excess of recombinant human IL-2 Cat. No.554603; data not shown , and by preincubation of the fixed/permeabilized cells with an excess of the unlabelled MQ1-17H12 antibody Cat. No. 554563; data not shown prior to staining with the FITC-MQ1-17H12 antibody. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the recombinant cytokine blocking and unlabelled antibody blocking specificity controls.