参考图片
Expression of IL-2 by stimulated CD4+ and CD4- BALB/c spleen cells. Splenocytes from 6 month old BALB/c mice were stimulated for 5 hours with hamster anti-mouse CD3 (2 µg/ml final concentration; clone 145-2C11, Cat. No. 553057) and hamster anti-mouse CD28 (2 µg/ml final concentration; clone 37.51, Cat. No. 553294) antibodies in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The splenocytes were harvested, stained with 0.06 µg of FITC-conjugated rat anti-mouse CD4 (FITC-RM4-5, Cat. No. 553047), fixed, permeabilized, and subsequently stained with 0.25 µg of PE anti-mouse IL-2 (Cat. No. 554428) (left panel). To demonstrate specificity, PE-JES6-5H4 staining was blocked by each of the following: 1) preincubation of the conjugated antibody with recombinant mouse IL-2 (0.25 µg, Cat. No. 550069; middle panel) and by 2) preincubation of the fixed/permeabilized cells with unlabelled JES6-5H4 mAb (5.0 µg; Cat. No. 554425) prior to staining with PE-JES6-5H4 (right panel). The quadrant markers for the bivariate dot plots were set based on the autofluorescence controls and verified using the recombinant cytokine blocking and unlabelled antibody blocking specificity controls.
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