BD Pharmingen- Purified Rat Anti-Human IL-2_BD Pharmingen
参考图片
Expression of IL-2 by stimulated CD3+ human PBMC. Human PBMCs were stimulated for 6 hours with PMA (50 ng/ml final concentration; Sigma, Cat. #P-8139) and calcium ionophore A23187 (500-100 ng/ml final concentration; Sigma, Cat. #C-9275) in the presence of BD GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMCs were stained with PE-Cy™5-anti-CD3 (Cat. No. 555334), fixed, permeabilized, and then stained with 0.25 µg of FITC-rat anti-human IL-2 antibody (Cat. No. 554565) using Pharmingen's staining protocol (left panel). To demonstrate specificity of staining, the binding of FITC-MQ1-17H12 was blocked by the preincubation of the fluorochrome - conjugated antibody with recombinant human IL-2 (10 ng/ml final concentration; Cat. No. 554603; middle panel), and by preincubation of the fixed/permeabilized cells with Purified Rat Anti-Human IL-2 (5.0 µg, Cat. No. 554563; right panel) 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 (middle panel) and unlabeled antibody blocking (right panel) specificity controls.
Expression of IL-2 by stimulated CD3+ human PBMC. Human PBMCs were stimulated for 6 hours with PMA (50 ng/ml final concentration; Sigma, Cat. #P-8139) and calcium ionophore A23187 (500-100 ng/ml final concentration; Sigma, Cat. #C-9275) in the presence of BD GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMCs were stained with PE-Cy™5-anti-CD3 (Cat. No. 555334), fixed, permeabilized, and then stained with 0.25 µg of FITC-rat anti-human IL-2 antibody (Cat. No. 554565) using Pharmingen's staining protocol (left panel). To demonstrate specificity of staining, the binding of FITC-MQ1-17H12 was blocked by the preincubation of the fluorochrome - conjugated antibody with recombinant human IL-2 (10 ng/ml final concentration; Cat. No. 554603; middle panel), and by preincubation of the fixed/permeabilized cells with Purified Rat Anti-Human IL-2 (5.0 µg, Cat. No. 554563; right panel) 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 (middle panel) and unlabeled antibody blocking (right panel) specificity controls.
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