BD Pharmingen- PE Rat Anti-Mouse-Anti-Human IL-5_BD Pharmingen
参考图片
Expression of IL-5 by stimulated mouse CD4+ T cells. Purified splenic CD4+ cells from 6-month old BALB/c mice were stimulated with plate-bound anti-CD3 (clone 145-2C11, Cat. No. 553057 at 25 µg/ml) and soluble anti-mouse CD28 (clone 37.51, Cat. No. 553294 at 2 µg/ml) for 2 days in culture together with recombinant mouse IL-2 (10 ng/ml, Cat. No. 550069) and recombinant mouse IL-4 (0.5 ng/ml, Cat. No. 550067), followed by a 3 day incubation with only recombinant IL-2 and IL-4. This was followed by a 5 hour stimulation with plate-bound anti-CD3 (25 µg/ml) and anti-mouse CD28 (2 µg/ml) in the presence of BD GolgiStop™ (Cat. No. 554724). The cells were harvested, stained with 0.05 µg of FITC-conjugated rat anti-mouse CD4 (FITC-RM4-5, Cat. No. 553047), fixed, permeabilized, and subsequently stained with 0.12 µg of PE-conjugated rat anti-mouse/human IL-5 antibody (PE-TRFK5, Cat. No. 554395) by using the BD Pharmingen staining protocol (left panel). The binding of PE-TRFK5 was blocked by preincubation of the conjugate with recombinant mouse IL-5 (0.25 µg; Cat. No. 554581; right panel). The quadrant markers for the bivariate dot plots were set based on the unstained cell control.
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