Characterization of IL-17A-producing cells within a stimulated mouse EL4 thymoma cell population. EL4 cells were cultured overnight in the presence of GolgiPlug™ (1 µg/ml final concentration; Cat. No. 555029) without (left panel) or with PMA (5 ng/ml final concentration; Sigma, Cat. No. P-8139) and Ionomycin (500 ng/ml final concentration; Sigma, Cat. No.I-0634) (middle and right panels). The cells were fixed, permeabilized and subsequently stained with 0.125 µg of PE conjugated anti-mouse IL-17A (PE-TC11-18H10, Cat. No.559502) using BD Pharmingen I/C staining protocol (left and middle panels) . To demonstrate the specificity of staining, the binding of PE-TC11-18H10 was blocked by the preincubation of the fixed/permeabilized, stimulated EL4 cells with unlabeled TC11-18H10 antibody (10 µg; right panel). The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the unstimulated stained (left panel) and unlabeled antibody blocking (right panel) controls.

Mouse IL-17A Standard Curve