BD PharmingenTM FITC Mouse Anti-Rat IFN-gamma

Description

Expression of IFN-gamma by stimulated LOU rat lymphoid cells. Lymphoid cells from LOU rat were stimulated for 2 days with plate bound purified NA/LE mouse anti-rat CD3 10 mug/ml; Cat. No. 554829 , purified NA/LE mouse anti-rat CD28 2 mug/ml; Cat. No.554993 , recombinant rat IL-2 10 ng/ml; Cat. No.555106 and recombinant rat IL-4 50 ng/ml; Cat. No. 555107 . The cells were then incubated for 3 days with recombinant rat IL-2 and IL-4. Following the 3 day incubation, the lymphoid cells were restimulated for 4 hours with PMA 5 ng/ml final concentration, Sigma, Cat. No.P-8139 and ionomycin 500 ng/ml final concentration, Sigma, Cat. No.I-0634 in the presence of GolgiPlug 1 mul/ml, Cat. No.555029 . The activated cells were harvested, fixed, permeabilized, and subsequently stained with 20 mul of FITC- conjugated mouse anti-rat IFN- antibody FITC-DB-1, Cat. No. 559498 by using Pharmingens staining protocol see Center panel . To demonstrate specificity of staining, the binding by the FITC-DB-1 antibody was blocked by pre-incubation of the fixed/permeabilized cells with unlabeled DB-1 antibody 5 mug; Right panel prior to staining. The quadrant markers for the bivariate dot plots were set based on the staining profile using FITC-MOPC-21 isotype control antibody Cat. No. 554679, see Left panel and verified using the unlabeled antibody blocking specificity control. Expression of IFN-gamma by stimulated LOU rat lymphoid cells. Lymphoid cells from LOU rat were stimulated for 2 days with plate bound purified NA/LE mouse anti-rat CD3 10 mug/ml; Cat. No. 554829 , purified NA/LE mouse anti-rat CD28 2 mug/ml; Cat. No.554993 , recombinant rat IL-2 10 ng/ml; Cat. No.555106 and recombinant rat IL-4 50 ng/ml; Cat. No. 555107 . The cells were then incubated for 3 days with recombinant rat IL-2 and IL-4. Following the 3 day incubation, the lymphoid cells were restimulated for 4 hours with PMA 5 ng/ml final concentration, Sigma, Cat. No.P-8139 and ionomycin 500 ng/ml final concentration, Sigma, Cat. No.I-0634 in the presence of GolgiPlug 1 mul/ml, Cat. No.555029 . The activated cells were harvested, fixed, permeabilized, and subsequently stained with 20 mul of FITC- conjugated mouse anti-rat IFN- antibody FITC-DB-1, Cat. No. 559498 by using Pharmingens staining protocol see Center panel . To demonstrate specificity of staining, the binding by the FITC-DB-1 antibody was blocked by pre-incubation of the fixed/permeabilized cells with unlabeled DB-1 antibody 5 mug; Right panel prior to staining. The quadrant markers for the bivariate dot plots were set based on the staining profile using FITC-MOPC-21 isotype control antibody Cat. No. 554679, see Left panel and verified using the unlabeled antibody blocking specificity control. Expression of IFN-gamma by stimulated LOU rat lymphoid cells. Lymphoid cells from LOU rat were stimulated for 2 days with plate bound purified NA/LE mouse anti-rat CD3 10 mug/ml; Cat. No. 554829 , purified NA/LE mouse anti-rat CD28 2 mug/ml; Cat. No.554993 , recombinant rat IL-2 10 ng/ml; Cat. No.555106 and recombinant rat IL-4 50 ng/ml; Cat. No. 555107 . The cells were then incubated for 3 days with recombinant rat IL-2 and IL-4. Following the 3 day incubation, the lymphoid cells were restimulated for 4 hours with PMA 5 ng/ml final concentration, Sigma, Cat. No.P-8139 and ionomycin 500 ng/ml final concentration, Sigma, Cat. No.I-0634 in the presence of GolgiPlug 1 mul/ml, Cat. No.555029 . The activated cells were harvested, fixed, permeabilized, and subsequently stained with 20 mul of FITC- conjugated mouse anti-rat IFN- antibody FITC-DB-1, Cat. No. 559498 by using Pharmingens staining protocol see Center panel . To demonstrate specificity of staining, the binding by the FITC-DB-1 antibody was blocked by pre-incubation of the fixed/permeabilized cells with unlabeled DB-1 antibody 5 mug; Right panel prior to staining. The quadrant markers for the bivariate dot plots were set based on the staining profile using FITC-MOPC-21 isotype control antibody Cat. No. 554679, see Left panel and verified using the unlabeled antibody blocking specificity control. Show More Expression of IFN-gamma by stimulated LOU rat lymphoid cells. Lymphoid cells from LOU rat were stimulated for 2 days with plate bound purified NA/LE mouse anti-rat CD3 10 mug/ml; Cat. No. 554829 , purified NA/LE mouse anti-rat CD28 2 mug/ml; Cat. No.554993 , recombinant rat IL-2 10 ng/ml; Cat. No.555106 and recombinant rat IL-4 50 ng/ml; Cat. No. 555107 . The cells were then incubated for 3 days with recombinant rat IL-2 and IL-4. Following the 3 day incubation, the lymphoid cells were restimulated for 4 hours with PMA 5 ng/ml final concentration, Sigma, Cat. No.P-8139 and ionomycin 500 ng/ml final concentration, Sigma, Cat. No.I-0634 in the presence of GolgiPlug 1 mul/ml, Cat. No.555029 . The activated cells were harvested, fixed, permeabilized, and subsequently stained with 20 mul of FITC- conjugated mouse anti-rat IFN- antibody FITC-DB-1, Cat. No. 559498 by using Pharmingens staining protocol see Center panel . To demonstrate specificity of staining, the binding by the FITC-DB-1 antibody was blocked by pre-incubation of the fixed/permeabilized cells with unlabeled DB-1 antibody 5 mug; Right panel prior to staining. The quadrant markers for the bivariate dot plots were set based on the staining profile using FITC-MOPC-21 isotype control antibody Cat. No. 554679, see Left panel and verified using the unlabeled antibody blocking specificity control. Show More Expression of IFN-gamma by stimulated LOU rat lymphoid cells. Lymphoid cells from LOU rat were stimulated for 2 days with plate bound purified NA/LE mouse anti-rat CD3 10 mug/ml; Cat. No. 554829 , purified NA/LE mouse anti-rat CD28 2 mug/ml; Cat. No.554993 , recombinant rat IL-2 10 ng/ml; Cat. No.555106 and recombinant rat IL-4 50 ng/ml; Cat. No. 555107 . The cells were then incubated for 3 days with recombinant rat IL-2 and IL-4. Following the 3 day incubation, the lymphoid cells were restimulated for 4 hours with PMA 5 ng/ml final concentration, Sigma, Cat. No.P-8139 and ionomycin 500 ng/ml final concentration, Sigma, Cat. No.I-0634 in the presence of GolgiPlug 1 mul/ml, Cat. No.555029 . The activated cells were harvested, fixed, permeabilized, and subsequently stained with 20 mul of FITC- conjugated mouse anti-rat IFN- antibody FITC-DB-1, Cat. No. 559498 by using Pharmingens staining protocol see Center panel . To demonstrate specificity of staining, the binding by the FITC-DB-1 antibody was blocked by pre-incubation of the fixed/permeabilized cells with unlabeled DB-1 antibody 5 mug; Right panel prior to staining. The quadrant markers for the bivariate dot plots were set based on the staining profile using FITC-MOPC-21 isotype control antibody Cat. No. 554679, see Left panel and verified using the unlabeled antibody blocking specificity control. Expression of IFN-gamma by stimulated LOU rat lymphoid cells. Lymphoid cells from LOU rat were stimulated for 2 days with plate bound purified NA/LE mouse anti-rat CD3 10 mug/ml; Cat. No. 554829 , purified NA/LE mouse anti-rat CD28 2 mug/ml; Cat. No.554993 , recombinant rat IL-2 10 ng/ml; Cat. No.555106 and recombinant rat IL-4 50 ng/ml; Cat. No. 555107 . The cells were then incubated for 3 days with recombinant rat IL-2 and IL-4. Following the 3 day incubation, the lymphoid cells were restimulated for 4 hours with PMA 5 ng/ml final concentration, Sigma, Cat. No.P-8139 and ionomycin 500 ng/ml final concentration, Sigma, Cat. No.I-0634 in the presence of GolgiPlug 1 mul/ml, Cat. No.555029 . The activated cells were harvested, fixed, permeabilized, and subsequently stained with 20 mul of FITC- conjugated mouse anti-rat IFN- antibody FITC-DB-1, Cat. No. 559498 by using Pharmingens staining protocol see Center panel . To demonstrate specificity of staining, the binding by the FITC-DB-1 antibody was blocked by pre-incubation of the fixed/permeabilized cells with unlabeled DB-1 antibody 5 mug; Right panel prior to staining. The quadrant markers for the bivariate dot plots were set based on the staining profile using FITC-MOPC-21 isotype control antibody Cat. No. 554679, see Left panel and verified using the unlabeled antibody blocking specificity control.