BD PharmingenTM PE-CyTM7 Rat Anti-Mouse TNF

Description

Expression of TNF by stimulated CD4+ and CD4- BALB/c spleen cells. Splenocytes from BALB/c mice were stimulated for 4 hours with PMA 5 ng/ml, Sigma, P-8139 and Ionomycin 500 ng, Sigma I-0634 in the presence of Brefeldin A GolgiPlug, Cat. No. 555029 . Cells were harvested, fixed, permeabilized and stained with APC-conjugated rat anti-mouse CD4 APC-RM4-5, Cat. No. 553051 and either rat anti-mouse TNF antibody PE-Cy7-MP6-XT22, Cat. No. 557644 left panel or immunoglobulin isotype control PE-Cy7-R3-34, Cat. No. 557645 , right panel by using BD Pharmingen staining protocol. To demonstrate specificity of staining the binding of PE-Cy7-MP6-XT22 was blocked by the preincubation of the conjugated antibody with molar excess of recombinant mouse TNF 0.25 mug, Cat. No. 554589, data not shown and by preincubation of the fixed/permeabilized cells with an excess of unlabelled MP6-XT22 antibody 5 mug, Cat. No. 554416, data not shown prior to staining. The quadrant markers for the bivariate dot plots were set based on the autofluorescence and isotype controls. Expression of TNF by stimulated CD4+ and CD4- BALB/c spleen cells. Splenocytes from BALB/c mice were stimulated for 4 hours with PMA 5 ng/ml, Sigma, P-8139 and Ionomycin 500 ng, Sigma I-0634 in the presence of Brefeldin A GolgiPlug, Cat. No. 555029 . Cells were harvested, fixed, permeabilized and stained with APC-conjugated rat anti-mouse CD4 APC-RM4-5, Cat. No. 553051 and either rat anti-mouse TNF antibody PE-Cy7-MP6-XT22, Cat. No. 557644 left panel or immunoglobulin isotype control PE-Cy7-R3-34, Cat. No. 557645 , right panel by using BD Pharmingen staining protocol. To demonstrate specificity of staining the binding of PE-Cy7-MP6-XT22 was blocked by the preincubation of the conjugated antibody with molar excess of recombinant mouse TNF 0.25 mug, Cat. No. 554589, data not shown and by preincubation of the fixed/permeabilized cells with an excess of unlabelled MP6-XT22 antibody 5 mug, Cat. No. 554416, data not shown prior to staining. The quadrant markers for the bivariate dot plots were set based on the autofluorescence and isotype controls. Expression of TNF by stimulated CD4+ and CD4- BALB/c spleen cells. Splenocytes from BALB/c mice were stimulated for 4 hours with PMA 5 ng/ml, Sigma, P-8139 and Ionomycin 500 ng, Sigma I-0634 in the presence of Brefeldin A GolgiPlug, Cat. No. 555029 . Cells were harvested, fixed, permeabilized and stained with APC-conjugated rat anti-mouse CD4 APC-RM4-5, Cat. No. 553051 and either rat anti-mouse TNF antibody PE-Cy7-MP6-XT22, Cat. No. 557644 left panel or immunoglobulin isotype control PE-Cy7-R3-34, Cat. No. 557645 , right panel by using BD Pharmingen staining protocol. To demonstrate specificity of staining the binding of PE-Cy7-MP6-XT22 was blocked by the preincubation of the conjugated antibody with molar excess of recombinant mouse TNF 0.25 mug, Cat. No. 554589, data not shown and by preincubation of the fixed/permeabilized cells with an excess of unlabelled MP6-XT22 antibody 5 mug, Cat. No. 554416, data not shown prior to staining. The quadrant markers for the bivariate dot plots were set based on the autofluorescence and isotype controls. Show More Expression of TNF by stimulated CD4+ and CD4- BALB/c spleen cells. Splenocytes from BALB/c mice were stimulated for 4 hours with PMA 5 ng/ml, Sigma, P-8139 and Ionomycin 500 ng, Sigma I-0634 in the presence of Brefeldin A GolgiPlug, Cat. No. 555029 . Cells were harvested, fixed, permeabilized and stained with APC-conjugated rat anti-mouse CD4 APC-RM4-5, Cat. No. 553051 and either rat anti-mouse TNF antibody PE-Cy7-MP6-XT22, Cat. No. 557644 left panel or immunoglobulin isotype control PE-Cy7-R3-34, Cat. No. 557645 , right panel by using BD Pharmingen staining protocol. To demonstrate specificity of staining the binding of PE-Cy7-MP6-XT22 was blocked by the preincubation of the conjugated antibody with molar excess of recombinant mouse TNF 0.25 mug, Cat. No. 554589, data not shown and by preincubation of the fixed/permeabilized cells with an excess of unlabelled MP6-XT22 antibody 5 mug, Cat. No. 554416, data not shown prior to staining. The quadrant markers for the bivariate dot plots were set based on the autofluorescence and isotype controls. Show More Expression of TNF by stimulated CD4+ and CD4- BALB/c spleen cells. Splenocytes from BALB/c mice were stimulated for 4 hours with PMA 5 ng/ml, Sigma, P-8139 and Ionomycin 500 ng, Sigma I-0634 in the presence of Brefeldin A GolgiPlug, Cat. No. 555029 . Cells were harvested, fixed, permeabilized and stained with APC-conjugated rat anti-mouse CD4 APC-RM4-5, Cat. No. 553051 and either rat anti-mouse TNF antibody PE-Cy7-MP6-XT22, Cat. No. 557644 left panel or immunoglobulin isotype control PE-Cy7-R3-34, Cat. No. 557645 , right panel by using BD Pharmingen staining protocol. To demonstrate specificity of staining the binding of PE-Cy7-MP6-XT22 was blocked by the preincubation of the conjugated antibody with molar excess of recombinant mouse TNF 0.25 mug, Cat. No. 554589, data not shown and by preincubation of the fixed/permeabilized cells with an excess of unlabelled MP6-XT22 antibody 5 mug, Cat. No. 554416, data not shown prior to staining. The quadrant markers for the bivariate dot plots were set based on the autofluorescence and isotype controls. Expression of TNF by stimulated CD4+ and CD4- BALB/c spleen cells. Splenocytes from BALB/c mice were stimulated for 4 hours with PMA 5 ng/ml, Sigma, P-8139 and Ionomycin 500 ng, Sigma I-0634 in the presence of Brefeldin A GolgiPlug, Cat. No. 555029 . Cells were harvested, fixed, permeabilized and stained with APC-conjugated rat anti-mouse CD4 APC-RM4-5, Cat. No. 553051 and either rat anti-mouse TNF antibody PE-Cy7-MP6-XT22, Cat. No. 557644 left panel or immunoglobulin isotype control PE-Cy7-R3-34, Cat. No. 557645 , right panel by using BD Pharmingen staining protocol. To demonstrate specificity of staining the binding of PE-Cy7-MP6-XT22 was blocked by the preincubation of the conjugated antibody with molar excess of recombinant mouse TNF 0.25 mug, Cat. No. 554589, data not shown and by preincubation of the fixed/permeabilized cells with an excess of unlabelled MP6-XT22 antibody 5 mug, Cat. No. 554416, data not shown prior to staining. The quadrant markers for the bivariate dot plots were set based on the autofluorescence and isotype controls.