BD PharmingenTM APC Rat Anti-Mouse TNF

Description

Expression of TNF by stimulated BALB/c spleen cells. After a 4 hour stimulation with PMA 5.0 ng/ml final concentration; Sigma and ionomycin 500 ng/ml final concentration; Sigma in the presence of GolgiPlug 1.0 mug/ml final concentration; Cat. No. 555029 the splenocytes were stained with FcBlock 1.0 mug/1 million cells; Cat No. 553142 . The cells were then fixed, permeabilized, and subsequently stained with 0.03 mug of APC-conjugated rat anti-mouse TNF antibody APC-MP6-XT22, Cat. No. 554420 by using Pharmingens staining protocol see Figure, left panel . To demonstrate specificity of staining, the binding of the APC-MP6-XT22 antibody was blocked by preincubation of the antibody conjugate with recombinant mouse TNF 0.25 mug, Cat. No. 554589; see middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled MP6-XT22 antibody 5.0 mug, Cat. No. 554416; see right panel prior to staining. 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 antibody blocking right panel specificity controls. This APC-conjugated reagent can be used in any flow cytometer equipped with a a dye, HeNE or red diode laser. These include the dual laser FACStarPLUS, FACS Vantage or FACSCalibur. Expression of TNF by stimulated BALB/c spleen cells. After a 4 hour stimulation with PMA 5.0 ng/ml final concentration; Sigma and ionomycin 500 ng/ml final concentration; Sigma in the presence of GolgiPlug 1.0 mug/ml final concentration; Cat. No. 555029 the splenocytes were stained with FcBlock 1.0 mug/1 million cells; Cat No. 553142 . The cells were then fixed, permeabilized, and subsequently stained with 0.03 mug of APC-conjugated rat anti-mouse TNF antibody APC-MP6-XT22, Cat. No. 554420 by using Pharmingens staining protocol see Figure, left panel . To demonstrate specificity of staining, the binding of the APC-MP6-XT22 antibody was blocked by preincubation of the antibody conjugate with recombinant mouse TNF 0.25 mug, Cat. No. 554589; see middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled MP6-XT22 antibody 5.0 mug, Cat. No. 554416; see right panel prior to staining. 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 antibody blocking right panel specificity controls. This APC-conjugated reagent can be used in any flow cytometer equipped with a a dye, HeNE or red diode laser. These include the dual laser FACStarPLUS, FACS Vantage or FACSCalibur. Expression of TNF by stimulated BALB/c spleen cells. After a 4 hour stimulation with PMA 5.0 ng/ml final concentration; Sigma and ionomycin 500 ng/ml final concentration; Sigma in the presence of GolgiPlug 1.0 mug/ml final concentration; Cat. No. 555029 the splenocytes were stained with FcBlock 1.0 mug/1 million cells; Cat No. 553142 . The cells were then fixed, permeabilized, and subsequently stained with 0.03 mug of APC-conjugated rat anti-mouse TNF antibody APC-MP6-XT22, Cat. No. 554420 by using Pharmingens staining protocol see Figure, left panel . To demonstrate specificity of staining, the binding of the APC-MP6-XT22 antibody was blocked by preincubation of the antibody conjugate with recombinant mouse TNF 0.25 mug, Cat. No. 554589; see middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled MP6-XT22 antibody 5.0 mug, Cat. No. 554416; see right panel prior to staining. 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 antibody blocking right panel specificity controls. This APC-conjugated reagent can be used in any flow cytometer equipped with a a dye, HeNE or red diode laser. These include the dual laser FACStarPLUS, FACS Vantage or FACSCalibur. Show More Expression of TNF by stimulated BALB/c spleen cells. After a 4 hour stimulation with PMA 5.0 ng/ml final concentration; Sigma and ionomycin 500 ng/ml final concentration; Sigma in the presence of GolgiPlug 1.0 mug/ml final concentration; Cat. No. 555029 the splenocytes were stained with FcBlock 1.0 mug/1 million cells; Cat No. 553142 . The cells were then fixed, permeabilized, and subsequently stained with 0.03 mug of APC-conjugated rat anti-mouse TNF antibody APC-MP6-XT22, Cat. No. 554420 by using Pharmingens staining protocol see Figure, left panel . To demonstrate specificity of staining, the binding of the APC-MP6-XT22 antibody was blocked by preincubation of the antibody conjugate with recombinant mouse TNF 0.25 mug, Cat. No. 554589; see middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled MP6-XT22 antibody 5.0 mug, Cat. No. 554416; see right panel prior to staining. 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 antibody blocking right panel specificity controls. This APC-conjugated reagent can be used in any flow cytometer equipped with a a dye, HeNE or red diode laser. These include the dual laser FACStarPLUS, FACS Vantage or FACSCalibur. Show More Expression of TNF by stimulated BALB/c spleen cells. After a 4 hour stimulation with PMA 5.0 ng/ml final concentration; Sigma and ionomycin 500 ng/ml final concentration; Sigma in the presence of GolgiPlug 1.0 mug/ml final concentration; Cat. No. 555029 the splenocytes were stained with FcBlock 1.0 mug/1 million cells; Cat No. 553142 . The cells were then fixed, permeabilized, and subsequently stained with 0.03 mug of APC-conjugated rat anti-mouse TNF antibody APC-MP6-XT22, Cat. No. 554420 by using Pharmingens staining protocol see Figure, left panel . To demonstrate specificity of staining, the binding of the APC-MP6-XT22 antibody was blocked by preincubation of the antibody conjugate with recombinant mouse TNF 0.25 mug, Cat. No. 554589; see middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled MP6-XT22 antibody 5.0 mug, Cat. No. 554416; see right panel prior to staining. 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 antibody blocking right panel specificity controls. This APC-conjugated reagent can be used in any flow cytometer equipped with a a dye, HeNE or red diode laser. These include the dual laser FACStarPLUS, FACS Vantage or FACSCalibur. Expression of TNF by stimulated BALB/c spleen cells. After a 4 hour stimulation with PMA 5.0 ng/ml final concentration; Sigma and ionomycin 500 ng/ml final concentration; Sigma in the presence of GolgiPlug 1.0 mug/ml final concentration; Cat. No. 555029 the splenocytes were stained with FcBlock 1.0 mug/1 million cells; Cat No. 553142 . The cells were then fixed, permeabilized, and subsequently stained with 0.03 mug of APC-conjugated rat anti-mouse TNF antibody APC-MP6-XT22, Cat. No. 554420 by using Pharmingens staining protocol see Figure, left panel . To demonstrate specificity of staining, the binding of the APC-MP6-XT22 antibody was blocked by preincubation of the antibody conjugate with recombinant mouse TNF 0.25 mug, Cat. No. 554589; see middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled MP6-XT22 antibody 5.0 mug, Cat. No. 554416; see right panel prior to staining. 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 antibody blocking right panel specificity controls. This APC-conjugated reagent can be used in any flow cytometer equipped with a a dye, HeNE or red diode laser. These include the dual laser FACStarPLUS, FACS Vantage or FACSCalibur.