BD PharmingenTM APC Mouse Anti-Human IL-12 (p40/p70)

Description

Expression of IL-12 p40/p70 by activated CD14+ human PBMCs. Ficoll-separated human PBMCs were primed for 2 hours with recombinant human IFN-gamma 10 ng/ml final concentration; Cat. No. 554616 , then activated with IFN-gamma 10 ng/ml final concentration and LPS 100 ng/ml final concentration; Sigma in the presence of GolgiStop 2 muM 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 mug of APC-C11.5 antibody Cat. No. 554576 , following Pharmingens 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 mug 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-gamma 10 ng/ml final concentration; Cat. No. 554616 , then activated with IFN-gamma 10 ng/ml final concentration and LPS 100 ng/ml final concentration; Sigma in the presence of GolgiStop 2 muM 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 mug of APC-C11.5 antibody Cat. No. 554576 , following Pharmingens 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 mug 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. Show More Expression of IL-12 p40/p70 by activated CD14+ human PBMCs. Ficoll-separated human PBMCs were primed for 2 hours with recombinant human IFN-gamma 10 ng/ml final concentration; Cat. No. 554616 , then activated with IFN-gamma 10 ng/ml final concentration and LPS 100 ng/ml final concentration; Sigma in the presence of GolgiStop 2 muM 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 mug of APC-C11.5 antibody Cat. No. 554576 , following Pharmingens 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 mug 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.