BD Pharmingen- APC Rat Anti-Human IL-10_BD Pharmingen
参考图片
Expression of IL-10 by stimulated CD14+ human monocytes. Human PBMC were stimulated for 24 hours with LPS (1 µg/ml final concentration) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were harvested, stained with FITC-mouse anti-human CD14 antibody (FITC-M5E2, Cat. No. 555397), fixed, permeabilized, and subsequently stained with 0.007 µg of APC-rat anti-human IL-10 antibody (APC-JES3-19F1, Cat. No. 554707) following the BD Bioscience staining protocol (left panel). The data reflect gating on monocytes, based on forward and side scatter. To demonstrate specificity of staining, the binding of APC-JES3-19F1 was blocked by the preincubation of the conjugated antibody with recombinant human IL-10 (0.25 µg, Cat. No. 554611; middle panel), and by preincubation of the fixed/permeabilized cells with unlabelled JES3-19F1 antibody (10 µg, Cat. No. 554705; right panel) prior to staining with the APC-JES3-19F1 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 unlabelled antibody (right panel) blocking specificity controls. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNE or red diode laser. These include the dual laser FACStarPLUS™, FACS Vantage™ or FACSCalibur™.
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