参考图片
Expression of IL-2 by stimulated CD3+ and CD3-human PBMC. Human PBMC were stimulated for 18 hours with PMA (Sigma, Cat. No. P-8139) and ionomycin (Sigma, Cat. No. I-0634), in the presence of BD GolgiStop™ (2 μl final concentration; Cat. No. 554724). The PBMC were stained with PE-Cy™5-anti-CD3 (Cat. No. 555334), fixed, permeabilized, and then stained with 20 μl of the PE Rat Anti-Human IL-2 antibody (Cat. No. 560902/559334/554566; left panel). To demonstrate specificity of staining, the binding of PE-MQ1-17H12 was blocked by the preincubation of the conjugated antibody with recombinant human IL-2 (0.25 mg, Cat. No. 554603; middle panel), and by preincubation of the fixed/permeabilized cells with an excess of Purified Rat Anti-Human IL-2 (10 mg, Cat. No. 554563; right panel) prior to staining with the PE-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 specificity controls (right panel).
Expression of IL-2 by stimulated CD3+ and CD3-human PBMC. Human PBMC were stimulated for 18 hours with PMA (Sigma, Cat. No. P-8139) and ionomycin (Sigma, Cat. No. I-0634), in the presence of BD GolgiStop™ (2 μl final concentration; Cat. No. 554724). The PBMC were stained with PE-Cy™5-anti-CD3 (Cat. No. 555334), fixed, permeabilized, and then stained with 20 μl of the PE Rat Anti-Human IL-2 antibody (Cat. No. 560902/559334/554566; left panel). To demonstrate specificity of staining, the binding of PE-MQ1-17H12 was blocked by the preincubation of the conjugated antibody with recombinant human IL-2 (0.25 mg, Cat. No. 554603; middle panel), and by preincubation of the fixed/permeabilized cells with an excess of Purified Rat Anti-Human IL-2 (10 mg, Cat. No. 554563; right panel) prior to staining with the PE-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 specificity controls (right panel).
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