BD PharmingenTM Purified Rat Anti-Mouse TNF

Description

Expression of TNF by stimulated CD4+ and CD4- BALB/c spleen cells. After 6 hour stimulation with hamster anti-mouse CD3 2 mug/ml final concentration; Clone 145-2C11, Cat. No. 553057 and hamster anti-mouse CD28 2 mug/ml final concentration; Clone 37.51, Cat. No. 553294 antibodies in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 , the splenocytes were stained with FcBlock 1 mug/1 million cells; Cat No. 553142 , then 0.06 mug of PE-conjugated rat anti-mouse CD4 PE-RM4-5, Cat. No. 553049 . The cells were then fixed, permeabilized, and subsequently stained with 0.06 mug of FITC-conjugated rat anti-mouse TNF antibody FITC-MP6-XT22, Cat. No. 554418 by using Pharmingens staining protocol left panel . To demonstrate specificity of staining, the binding of the FITC-MP6-XT22 antibody was blocked by preincubation of the antibody conjugate with recombinant mouse TNF 0.25 mug, Cat. No. 554589; middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled MP6-XT22 antibody 2 mug, Cat.No. 554416; 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. Expression of TNF by stimulated CD4+ and CD4- BALB/c spleen cells. After 6 hour stimulation with hamster anti-mouse CD3 2 mug/ml final concentration; Clone 145-2C11, Cat. No. 553057 and hamster anti-mouse CD28 2 mug/ml final concentration; Clone 37.51, Cat. No. 553294 antibodies in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 , the splenocytes were stained with FcBlock 1 mug/1 million cells; Cat No. 553142 , then 0.06 mug of PE-conjugated rat anti-mouse CD4 PE-RM4-5, Cat. No. 553049 . The cells were then fixed, permeabilized, and subsequently stained with 0.06 mug of FITC-conjugated rat anti-mouse TNF antibody FITC-MP6-XT22, Cat. No. 554418 by using Pharmingens staining protocol left panel . To demonstrate specificity of staining, the binding of the FITC-MP6-XT22 antibody was blocked by preincubation of the antibody conjugate with recombinant mouse TNF 0.25 mug, Cat. No. 554589; middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled MP6-XT22 antibody 2 mug, Cat.No. 554416; 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. Expression of TNF by stimulated CD4+ and CD4- BALB/c spleen cells. After 6 hour stimulation with hamster anti-mouse CD3 2 mug/ml final concentration; Clone 145-2C11, Cat. No. 553057 and hamster anti-mouse CD28 2 mug/ml final concentration; Clone 37.51, Cat. No. 553294 antibodies in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 , the splenocytes were stained with FcBlock 1 mug/1 million cells; Cat No. 553142 , then 0.06 mug of PE-conjugated rat anti-mouse CD4 PE-RM4-5, Cat. No. 553049 . The cells were then fixed, permeabilized, and subsequently stained with 0.06 mug of FITC-conjugated rat anti-mouse TNF antibody FITC-MP6-XT22, Cat. No. 554418 by using Pharmingens staining protocol left panel . To demonstrate specificity of staining, the binding of the FITC-MP6-XT22 antibody was blocked by preincubation of the antibody conjugate with recombinant mouse TNF 0.25 mug, Cat. No. 554589; middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled MP6-XT22 antibody 2 mug, Cat.No. 554416; 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. Show More Expression of TNF by stimulated CD4+ and CD4- BALB/c spleen cells. After 6 hour stimulation with hamster anti-mouse CD3 2 mug/ml final concentration; Clone 145-2C11, Cat. No. 553057 and hamster anti-mouse CD28 2 mug/ml final concentration; Clone 37.51, Cat. No. 553294 antibodies in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 , the splenocytes were stained with FcBlock 1 mug/1 million cells; Cat No. 553142 , then 0.06 mug of PE-conjugated rat anti-mouse CD4 PE-RM4-5, Cat. No. 553049 . The cells were then fixed, permeabilized, and subsequently stained with 0.06 mug of FITC-conjugated rat anti-mouse TNF antibody FITC-MP6-XT22, Cat. No. 554418 by using Pharmingens staining protocol left panel . To demonstrate specificity of staining, the binding of the FITC-MP6-XT22 antibody was blocked by preincubation of the antibody conjugate with recombinant mouse TNF 0.25 mug, Cat. No. 554589; middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled MP6-XT22 antibody 2 mug, Cat.No. 554416; 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. Show More Expression of TNF by stimulated CD4+ and CD4- BALB/c spleen cells. After 6 hour stimulation with hamster anti-mouse CD3 2 mug/ml final concentration; Clone 145-2C11, Cat. No. 553057 and hamster anti-mouse CD28 2 mug/ml final concentration; Clone 37.51, Cat. No. 553294 antibodies in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 , the splenocytes were stained with FcBlock 1 mug/1 million cells; Cat No. 553142 , then 0.06 mug of PE-conjugated rat anti-mouse CD4 PE-RM4-5, Cat. No. 553049 . The cells were then fixed, permeabilized, and subsequently stained with 0.06 mug of FITC-conjugated rat anti-mouse TNF antibody FITC-MP6-XT22, Cat. No. 554418 by using Pharmingens staining protocol left panel . To demonstrate specificity of staining, the binding of the FITC-MP6-XT22 antibody was blocked by preincubation of the antibody conjugate with recombinant mouse TNF 0.25 mug, Cat. No. 554589; middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled MP6-XT22 antibody 2 mug, Cat.No. 554416; 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. Expression of TNF by stimulated CD4+ and CD4- BALB/c spleen cells. After 6 hour stimulation with hamster anti-mouse CD3 2 mug/ml final concentration; Clone 145-2C11, Cat. No. 553057 and hamster anti-mouse CD28 2 mug/ml final concentration; Clone 37.51, Cat. No. 553294 antibodies in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 , the splenocytes were stained with FcBlock 1 mug/1 million cells; Cat No. 553142 , then 0.06 mug of PE-conjugated rat anti-mouse CD4 PE-RM4-5, Cat. No. 553049 . The cells were then fixed, permeabilized, and subsequently stained with 0.06 mug of FITC-conjugated rat anti-mouse TNF antibody FITC-MP6-XT22, Cat. No. 554418 by using Pharmingens staining protocol left panel . To demonstrate specificity of staining, the binding of the FITC-MP6-XT22 antibody was blocked by preincubation of the antibody conjugate with recombinant mouse TNF 0.25 mug, Cat. No. 554589; middle panel , and by preincubation of the fixed/permeabilized cells with unlabeled MP6-XT22 antibody 2 mug, Cat.No. 554416; 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.