BD PharmingenTM PE Rat Anti-Mouse IL-3

Description

Expression of IL-3 by stimulated CD4+ Balb/c spleen cells. Purified splenic CD4+ cells from 6 month old BALB/c mice were stimulated with plate-bound anti-CD3 25 mug/ml final concentration; 145-2C11, Cat. No. 553057 and soluble anti-mouse CD28 2 mug /ml final concentration; clone 37.51, Cat. No. 553294 for 2 days in culture together with recombinant mouse IL-2 10 ng/ml final concentration; Cat. No. 550069 and recombinant mouse IL-4 0.5 ng/ml final concentration; Cat. No. 550067 , followed by a 3 day incubation with only recombinant mouse IL-2 and recombinant mouse IL-4. This was followed by a 5 hour stimulation with plate-bound anti-CD3 25 mug/ml final concentration and anti-mouse CD28 2 mug/ml final concentration in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 . The cells 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 0.12 mug of PE-conjugated rat anti-mouse IL-3 antibody PE-MP2-8F8, Cat. No. 554383 by using the BD Pharmingen staining protocol left panel . To demonstrate specificity of staining, the binding by PE-MP2-8F8 was blocked by each of the following: 1 preincubation of the conjugated antibody with molar excess of recombinant mouse IL-3 0.12 mug, Cat. No. 554579; center panel and by 2 preincubation of the fixed/permeabilized cells with excess unlabeled MP2-8F8 mouse antibody 3 mug; Cat. No. 554380; right panel prior to staining with the PE-MP2-8F8. The quadrant markers for the bivariate dot plots were set based on the autofluorescence controls and verified using the recombinant cytokine blocking and unlabeled antibody blocking specificity controls. Expression of IL-3 by stimulated CD4+ Balb/c spleen cells. Purified splenic CD4+ cells from 6 month old BALB/c mice were stimulated with plate-bound anti-CD3 25 mug/ml final concentration; 145-2C11, Cat. No. 553057 and soluble anti-mouse CD28 2 mug /ml final concentration; clone 37.51, Cat. No. 553294 for 2 days in culture together with recombinant mouse IL-2 10 ng/ml final concentration; Cat. No. 550069 and recombinant mouse IL-4 0.5 ng/ml final concentration; Cat. No. 550067 , followed by a 3 day incubation with only recombinant mouse IL-2 and recombinant mouse IL-4. This was followed by a 5 hour stimulation with plate-bound anti-CD3 25 mug/ml final concentration and anti-mouse CD28 2 mug/ml final concentration in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 . The cells 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 0.12 mug of PE-conjugated rat anti-mouse IL-3 antibody PE-MP2-8F8, Cat. No. 554383 by using the BD Pharmingen staining protocol left panel . To demonstrate specificity of staining, the binding by PE-MP2-8F8 was blocked by each of the following: 1 preincubation of the conjugated antibody with molar excess of recombinant mouse IL-3 0.12 mug, Cat. No. 554579; center panel and by 2 preincubation of the fixed/permeabilized cells with excess unlabeled MP2-8F8 mouse antibody 3 mug; Cat. No. 554380; right panel prior to staining with the PE-MP2-8F8. The quadrant markers for the bivariate dot plots were set based on the autofluorescence controls and verified using the recombinant cytokine blocking and unlabeled antibody blocking specificity controls. Expression of IL-3 by stimulated CD4+ Balb/c spleen cells. Purified splenic CD4+ cells from 6 month old BALB/c mice were stimulated with plate-bound anti-CD3 25 mug/ml final concentration; 145-2C11, Cat. No. 553057 and soluble anti-mouse CD28 2 mug /ml final concentration; clone 37.51, Cat. No. 553294 for 2 days in culture together with recombinant mouse IL-2 10 ng/ml final concentration; Cat. No. 550069 and recombinant mouse IL-4 0.5 ng/ml final concentration; Cat. No. 550067 , followed by a 3 day incubation with only recombinant mouse IL-2 and recombinant mouse IL-4. This was followed by a 5 hour stimulation with plate-bound anti-CD3 25 mug/ml final concentration and anti-mouse CD28 2 mug/ml final concentration in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 . The cells 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 0.12 mug of PE-conjugated rat anti-mouse IL-3 antibody PE-MP2-8F8, Cat. No. 554383 by using the BD Pharmingen staining protocol left panel . To demonstrate specificity of staining, the binding by PE-MP2-8F8 was blocked by each of the following: 1 preincubation of the conjugated antibody with molar excess of recombinant mouse IL-3 0.12 mug, Cat. No. 554579; center panel and by 2 preincubation of the fixed/permeabilized cells with excess unlabeled MP2-8F8 mouse antibody 3 mug; Cat. No. 554380; right panel prior to staining with the PE-MP2-8F8. The quadrant markers for the bivariate dot plots were set based on the autofluorescence controls and verified using the recombinant cytokine blocking and unlabeled antibody blocking specificity controls. Show More Expression of IL-3 by stimulated CD4+ Balb/c spleen cells. Purified splenic CD4+ cells from 6 month old BALB/c mice were stimulated with plate-bound anti-CD3 25 mug/ml final concentration; 145-2C11, Cat. No. 553057 and soluble anti-mouse CD28 2 mug /ml final concentration; clone 37.51, Cat. No. 553294 for 2 days in culture together with recombinant mouse IL-2 10 ng/ml final concentration; Cat. No. 550069 and recombinant mouse IL-4 0.5 ng/ml final concentration; Cat. No. 550067 , followed by a 3 day incubation with only recombinant mouse IL-2 and recombinant mouse IL-4. This was followed by a 5 hour stimulation with plate-bound anti-CD3 25 mug/ml final concentration and anti-mouse CD28 2 mug/ml final concentration in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 . The cells 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 0.12 mug of PE-conjugated rat anti-mouse IL-3 antibody PE-MP2-8F8, Cat. No. 554383 by using the BD Pharmingen staining protocol left panel . To demonstrate specificity of staining, the binding by PE-MP2-8F8 was blocked by each of the following: 1 preincubation of the conjugated antibody with molar excess of recombinant mouse IL-3 0.12 mug, Cat. No. 554579; center panel and by 2 preincubation of the fixed/permeabilized cells with excess unlabeled MP2-8F8 mouse antibody 3 mug; Cat. No. 554380; right panel prior to staining with the PE-MP2-8F8. The quadrant markers for the bivariate dot plots were set based on the autofluorescence controls and verified using the recombinant cytokine blocking and unlabeled antibody blocking specificity controls. Show More Expression of IL-3 by stimulated CD4+ Balb/c spleen cells. Purified splenic CD4+ cells from 6 month old BALB/c mice were stimulated with plate-bound anti-CD3 25 mug/ml final concentration; 145-2C11, Cat. No. 553057 and soluble anti-mouse CD28 2 mug /ml final concentration; clone 37.51, Cat. No. 553294 for 2 days in culture together with recombinant mouse IL-2 10 ng/ml final concentration; Cat. No. 550069 and recombinant mouse IL-4 0.5 ng/ml final concentration; Cat. No. 550067 , followed by a 3 day incubation with only recombinant mouse IL-2 and recombinant mouse IL-4. This was followed by a 5 hour stimulation with plate-bound anti-CD3 25 mug/ml final concentration and anti-mouse CD28 2 mug/ml final concentration in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 . The cells 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 0.12 mug of PE-conjugated rat anti-mouse IL-3 antibody PE-MP2-8F8, Cat. No. 554383 by using the BD Pharmingen staining protocol left panel . To demonstrate specificity of staining, the binding by PE-MP2-8F8 was blocked by each of the following: 1 preincubation of the conjugated antibody with molar excess of recombinant mouse IL-3 0.12 mug, Cat. No. 554579; center panel and by 2 preincubation of the fixed/permeabilized cells with excess unlabeled MP2-8F8 mouse antibody 3 mug; Cat. No. 554380; right panel prior to staining with the PE-MP2-8F8. The quadrant markers for the bivariate dot plots were set based on the autofluorescence controls and verified using the recombinant cytokine blocking and unlabeled antibody blocking specificity controls. Expression of IL-3 by stimulated CD4+ Balb/c spleen cells. Purified splenic CD4+ cells from 6 month old BALB/c mice were stimulated with plate-bound anti-CD3 25 mug/ml final concentration; 145-2C11, Cat. No. 553057 and soluble anti-mouse CD28 2 mug /ml final concentration; clone 37.51, Cat. No. 553294 for 2 days in culture together with recombinant mouse IL-2 10 ng/ml final concentration; Cat. No. 550069 and recombinant mouse IL-4 0.5 ng/ml final concentration; Cat. No. 550067 , followed by a 3 day incubation with only recombinant mouse IL-2 and recombinant mouse IL-4. This was followed by a 5 hour stimulation with plate-bound anti-CD3 25 mug/ml final concentration and anti-mouse CD28 2 mug/ml final concentration in the presence of GolgiStop 2 muM final concentration; Cat. No. 554724 . The cells 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 0.12 mug of PE-conjugated rat anti-mouse IL-3 antibody PE-MP2-8F8, Cat. No. 554383 by using the BD Pharmingen staining protocol left panel . To demonstrate specificity of staining, the binding by PE-MP2-8F8 was blocked by each of the following: 1 preincubation of the conjugated antibody with molar excess of recombinant mouse IL-3 0.12 mug, Cat. No. 554579; center panel and by 2 preincubation of the fixed/permeabilized cells with excess unlabeled MP2-8F8 mouse antibody 3 mug; Cat. No. 554380; right panel prior to staining with the PE-MP2-8F8. The quadrant markers for the bivariate dot plots were set based on the autofluorescence controls and verified using the recombinant cytokine blocking and unlabeled antibody blocking specificity controls.