BD PharmingenTM PE Rat Anti-Mouse IL-4

Description

Expression of IL-4 by stimulated CD4+ and CD4-BALB/c spleen cells. Splenocytes from 6 month old BALB/c mice were cultured for 72 hours in medium containing Staphylococcus aureus enterotoxin B 2 mug/mL final concentration; Sigma, St. Louis, MO , recombinant mouse IL-2 10 U/mL final concentration; Cat. No. 550069 and recombinant mouse IL-4 2 ng/mL final concentration; Cat. No. 550067 . The cells were harvested and restimulated for 5 hours with anti-CD3 2 mug/mL final concentration; 145-2C11, Cat. No. 553057 and anti-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 harvested, stained with 0.06 mug of FITC-conjugated rat anti-mouse CD4 FITC-RM4-5, Cat. No. 553047 fixed, permeabilized, and subsequently stained with 0.25 mug of PE-conjugated rat anti-mouse IL-4 antibody PE-BVD4-1D11, Cat. No. 554389 by using the BD Pharmingen staining protocol see left panel . To demonstrate specificity of staining, the binding of PE-BVD4-1D11 was blocked by the preincubation of the conjugated antibody with recombinant mouse IL-4 0.25 mug, Cat. No. 550067; see middle panel , and by preincubation of the fixed/permeabilized cells with unlabelled BVD4-1D11 antibody 5 mug, Cat. No. 554386; see right panel . 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. Expression of IL-4 by stimulated CD4+ and CD4-BALB/c spleen cells. Splenocytes from 6 month old BALB/c mice were cultured for 72 hours in medium containing Staphylococcus aureus enterotoxin B 2 mug/mL final concentration; Sigma, St. Louis, MO , recombinant mouse IL-2 10 U/mL final concentration; Cat. No. 550069 and recombinant mouse IL-4 2 ng/mL final concentration; Cat. No. 550067 . The cells were harvested and restimulated for 5 hours with anti-CD3 2 mug/mL final concentration; 145-2C11, Cat. No. 553057 and anti-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 harvested, stained with 0.06 mug of FITC-conjugated rat anti-mouse CD4 FITC-RM4-5, Cat. No. 553047 fixed, permeabilized, and subsequently stained with 0.25 mug of PE-conjugated rat anti-mouse IL-4 antibody PE-BVD4-1D11, Cat. No. 554389 by using the BD Pharmingen staining protocol see left panel . To demonstrate specificity of staining, the binding of PE-BVD4-1D11 was blocked by the preincubation of the conjugated antibody with recombinant mouse IL-4 0.25 mug, Cat. No. 550067; see middle panel , and by preincubation of the fixed/permeabilized cells with unlabelled BVD4-1D11 antibody 5 mug, Cat. No. 554386; see right panel . 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. Expression of IL-4 by stimulated CD4+ and CD4-BALB/c spleen cells. Splenocytes from 6 month old BALB/c mice were cultured for 72 hours in medium containing Staphylococcus aureus enterotoxin B 2 mug/mL final concentration; Sigma, St. Louis, MO , recombinant mouse IL-2 10 U/mL final concentration; Cat. No. 550069 and recombinant mouse IL-4 2 ng/mL final concentration; Cat. No. 550067 . The cells were harvested and restimulated for 5 hours with anti-CD3 2 mug/mL final concentration; 145-2C11, Cat. No. 553057 and anti-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 harvested, stained with 0.06 mug of FITC-conjugated rat anti-mouse CD4 FITC-RM4-5, Cat. No. 553047 fixed, permeabilized, and subsequently stained with 0.25 mug of PE-conjugated rat anti-mouse IL-4 antibody PE-BVD4-1D11, Cat. No. 554389 by using the BD Pharmingen staining protocol see left panel . To demonstrate specificity of staining, the binding of PE-BVD4-1D11 was blocked by the preincubation of the conjugated antibody with recombinant mouse IL-4 0.25 mug, Cat. No. 550067; see middle panel , and by preincubation of the fixed/permeabilized cells with unlabelled BVD4-1D11 antibody 5 mug, Cat. No. 554386; see right panel . 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. Show More Expression of IL-4 by stimulated CD4+ and CD4-BALB/c spleen cells. Splenocytes from 6 month old BALB/c mice were cultured for 72 hours in medium containing Staphylococcus aureus enterotoxin B 2 mug/mL final concentration; Sigma, St. Louis, MO , recombinant mouse IL-2 10 U/mL final concentration; Cat. No. 550069 and recombinant mouse IL-4 2 ng/mL final concentration; Cat. No. 550067 . The cells were harvested and restimulated for 5 hours with anti-CD3 2 mug/mL final concentration; 145-2C11, Cat. No. 553057 and anti-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 harvested, stained with 0.06 mug of FITC-conjugated rat anti-mouse CD4 FITC-RM4-5, Cat. No. 553047 fixed, permeabilized, and subsequently stained with 0.25 mug of PE-conjugated rat anti-mouse IL-4 antibody PE-BVD4-1D11, Cat. No. 554389 by using the BD Pharmingen staining protocol see left panel . To demonstrate specificity of staining, the binding of PE-BVD4-1D11 was blocked by the preincubation of the conjugated antibody with recombinant mouse IL-4 0.25 mug, Cat. No. 550067; see middle panel , and by preincubation of the fixed/permeabilized cells with unlabelled BVD4-1D11 antibody 5 mug, Cat. No. 554386; see right panel . 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. Show More Expression of IL-4 by stimulated CD4+ and CD4-BALB/c spleen cells. Splenocytes from 6 month old BALB/c mice were cultured for 72 hours in medium containing Staphylococcus aureus enterotoxin B 2 mug/mL final concentration; Sigma, St. Louis, MO , recombinant mouse IL-2 10 U/mL final concentration; Cat. No. 550069 and recombinant mouse IL-4 2 ng/mL final concentration; Cat. No. 550067 . The cells were harvested and restimulated for 5 hours with anti-CD3 2 mug/mL final concentration; 145-2C11, Cat. No. 553057 and anti-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 harvested, stained with 0.06 mug of FITC-conjugated rat anti-mouse CD4 FITC-RM4-5, Cat. No. 553047 fixed, permeabilized, and subsequently stained with 0.25 mug of PE-conjugated rat anti-mouse IL-4 antibody PE-BVD4-1D11, Cat. No. 554389 by using the BD Pharmingen staining protocol see left panel . To demonstrate specificity of staining, the binding of PE-BVD4-1D11 was blocked by the preincubation of the conjugated antibody with recombinant mouse IL-4 0.25 mug, Cat. No. 550067; see middle panel , and by preincubation of the fixed/permeabilized cells with unlabelled BVD4-1D11 antibody 5 mug, Cat. No. 554386; see right panel . 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. Expression of IL-4 by stimulated CD4+ and CD4-BALB/c spleen cells. Splenocytes from 6 month old BALB/c mice were cultured for 72 hours in medium containing Staphylococcus aureus enterotoxin B 2 mug/mL final concentration; Sigma, St. Louis, MO , recombinant mouse IL-2 10 U/mL final concentration; Cat. No. 550069 and recombinant mouse IL-4 2 ng/mL final concentration; Cat. No. 550067 . The cells were harvested and restimulated for 5 hours with anti-CD3 2 mug/mL final concentration; 145-2C11, Cat. No. 553057 and anti-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 harvested, stained with 0.06 mug of FITC-conjugated rat anti-mouse CD4 FITC-RM4-5, Cat. No. 553047 fixed, permeabilized, and subsequently stained with 0.25 mug of PE-conjugated rat anti-mouse IL-4 antibody PE-BVD4-1D11, Cat. No. 554389 by using the BD Pharmingen staining protocol see left panel . To demonstrate specificity of staining, the binding of PE-BVD4-1D11 was blocked by the preincubation of the conjugated antibody with recombinant mouse IL-4 0.25 mug, Cat. No. 550067; see middle panel , and by preincubation of the fixed/permeabilized cells with unlabelled BVD4-1D11 antibody 5 mug, Cat. No. 554386; see right panel . 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.