BD Pharmingen- APC Mouse Anti-Human IL-12 -p40-p70_BD Pharmingen
参考图片
Expression of IL-12 p40/p70 by activated CD14+ human PBMCs. Ficoll™-separated human PBMCs were primed for 2 hours with recombinant human IFN-γ (10 ng/ml final concentration; Cat. No. 554616), then activated with IFN-γ (10 ng/ml final concentration) and LPS (100 ng/ml final concentration; Sigma) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724) for an additional 22 hours. Cells were harvested, stained with FITC-mouse anti human CD14 antibody (Cat. No. 555397), fixed, permeabilized, and then stained with 0.125 µg of APC-C11.5 antibody (Cat. No. 554576), following Pharmingen's staining protocol (far left panel). The data reflect gating on monocytes, based on forward and side scattered light signals. To demonstrate specificity of staining, the binding of APC-C11.5 antibody was blocked by preincubation of the conjugated antibody with excess recombinant human IL-12 p70 (Cat. No. 554613; middle left panel) and recombinant human IL-12 p40 (Cat. No. 554633; middle right panel), but not by recombinant human IL-12 p35 protein (far right panel). Preincubation of the fixed/permeabilized cells with an excess of the unlabelled C11.5 antibody (5 µg final; Cat. No. 554573) prior to staining with the APC-C11.5 antibody also blocked staining (data not shown). The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the recombinant cytokine blocking and unlabelled antibody 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.™
Expression of IL-12 p40/p70 by activated CD14+ human PBMCs. Ficoll™-separated human PBMCs were primed for 2 hours with recombinant human IFN-γ (10 ng/ml final concentration; Cat. No. 554616), then activated with IFN-γ (10 ng/ml final concentration) and LPS (100 ng/ml final concentration; Sigma) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724) for an additional 22 hours. Cells were harvested, stained with FITC-mouse anti human CD14 antibody (Cat. No. 555397), fixed, permeabilized, and then stained with 0.125 µg of APC-C11.5 antibody (Cat. No. 554576), following Pharmingen's staining protocol (far left panel). The data reflect gating on monocytes, based on forward and side scattered light signals. To demonstrate specificity of staining, the binding of APC-C11.5 antibody was blocked by preincubation of the conjugated antibody with excess recombinant human IL-12 p70 (Cat. No. 554613; middle left panel) and recombinant human IL-12 p40 (Cat. No. 554633; middle right panel), but not by recombinant human IL-12 p35 protein (far right panel). Preincubation of the fixed/permeabilized cells with an excess of the unlabelled C11.5 antibody (5 µg final; Cat. No. 554573) prior to staining with the APC-C11.5 antibody also blocked staining (data not shown). The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the recombinant cytokine blocking and unlabelled antibody 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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