BD PharmingenTM PE Mouse Anti-Human TNF

Description

Expression of TNF protein by stimulated human peripheral blood mononuclear cells PBMC . Human PBMC were stimulated for 6 hours with PMA 50 ng/ml final concentration; Sigma, Cat. #P-8139 , calcium ionophore A23187 0.5 mug/ml final concentration; Sigma, Cat. #C-9275 and PHA Gibco BRL, Cat. #10576-015 in the presence of GolgiStop 2 muM final concentration; Cat. No. 554715 . The PBMC were stained with 0.25 mug PE-Cy5 Mouse Anti-Human CD3 Cat. No. 555334 and subsequently stained with 20 mul of PE Mouse Anti-Human TNF Cat. No. 559321/554513/562083 by following the recommended assay procedure left panel . To demonstrate specificity of staining, the binding of PE-MAb11 was blocked by preincubation of the conjugate with excess 1 mug recombinant human TNF Cat. No. 554618; middle panel and by preincubation of the fixed/permeabilized cells with an excess of Purified Mouse Anti-Human TNF Cat. No. 554510; right panel prior to staining with the PE-MAb11 antibody. The quadrant markers for the bivariate dot plot was set based on the negative staining controls using isotype-matched Ig controls. Expression of TNF protein by stimulated human peripheral blood mononuclear cells PBMC . Human PBMC were stimulated for 6 hours with PMA 50 ng/ml final concentration; Sigma, Cat. #P-8139 , calcium ionophore A23187 0.5 mug/ml final concentration; Sigma, Cat. #C-9275 and PHA Gibco BRL, Cat. #10576-015 in the presence of GolgiStop 2 muM final concentration; Cat. No. 554715 . The PBMC were stained with 0.25 mug PE-Cy5 Mouse Anti-Human CD3 Cat. No. 555334 and subsequently stained with 20 mul of PE Mouse Anti-Human TNF Cat. No. 559321/554513/562083 by following the recommended assay procedure left panel . To demonstrate specificity of staining, the binding of PE-MAb11 was blocked by preincubation of the conjugate with excess 1 mug recombinant human TNF Cat. No. 554618; middle panel and by preincubation of the fixed/permeabilized cells with an excess of Purified Mouse Anti-Human TNF Cat. No. 554510; right panel prior to staining with the PE-MAb11 antibody. The quadrant markers for the bivariate dot plot was set based on the negative staining controls using isotype-matched Ig controls. Expression of TNF protein by stimulated human peripheral blood mononuclear cells PBMC . Human PBMC were stimulated for 6 hours with PMA 50 ng/ml final concentration; Sigma, Cat. #P-8139 , calcium ionophore A23187 0.5 mug/ml final concentration; Sigma, Cat. #C-9275 and PHA Gibco BRL, Cat. #10576-015 in the presence of GolgiStop 2 muM final concentration; Cat. No. 554715 . The PBMC were stained with 0.25 mug PE-Cy5 Mouse Anti-Human CD3 Cat. No. 555334 and subsequently stained with 20 mul of PE Mouse Anti-Human TNF Cat. No. 559321/554513/562083 by following the recommended assay procedure left panel . To demonstrate specificity of staining, the binding of PE-MAb11 was blocked by preincubation of the conjugate with excess 1 mug recombinant human TNF Cat. No. 554618; middle panel and by preincubation of the fixed/permeabilized cells with an excess of Purified Mouse Anti-Human TNF Cat. No. 554510; right panel prior to staining with the PE-MAb11 antibody. The quadrant markers for the bivariate dot plot was set based on the negative staining controls using isotype-matched Ig controls. Show More Expression of TNF protein by stimulated human peripheral blood mononuclear cells PBMC . Human PBMC were stimulated for 6 hours with PMA 50 ng/ml final concentration; Sigma, Cat. #P-8139 , calcium ionophore A23187 0.5 mug/ml final concentration; Sigma, Cat. #C-9275 and PHA Gibco BRL, Cat. #10576-015 in the presence of GolgiStop 2 muM final concentration; Cat. No. 554715 . The PBMC were stained with 0.25 mug PE-Cy5 Mouse Anti-Human CD3 Cat. No. 555334 and subsequently stained with 20 mul of PE Mouse Anti-Human TNF Cat. No. 559321/554513/562083 by following the recommended assay procedure left panel . To demonstrate specificity of staining, the binding of PE-MAb11 was blocked by preincubation of the conjugate with excess 1 mug recombinant human TNF Cat. No. 554618; middle panel and by preincubation of the fixed/permeabilized cells with an excess of Purified Mouse Anti-Human TNF Cat. No. 554510; right panel prior to staining with the PE-MAb11 antibody. The quadrant markers for the bivariate dot plot was set based on the negative staining controls using isotype-matched Ig controls. Show More Expression of TNF protein by stimulated human peripheral blood mononuclear cells PBMC . Human PBMC were stimulated for 6 hours with PMA 50 ng/ml final concentration; Sigma, Cat. #P-8139 , calcium ionophore A23187 0.5 mug/ml final concentration; Sigma, Cat. #C-9275 and PHA Gibco BRL, Cat. #10576-015 in the presence of GolgiStop 2 muM final concentration; Cat. No. 554715 . The PBMC were stained with 0.25 mug PE-Cy5 Mouse Anti-Human CD3 Cat. No. 555334 and subsequently stained with 20 mul of PE Mouse Anti-Human TNF Cat. No. 559321/554513/562083 by following the recommended assay procedure left panel . To demonstrate specificity of staining, the binding of PE-MAb11 was blocked by preincubation of the conjugate with excess 1 mug recombinant human TNF Cat. No. 554618; middle panel and by preincubation of the fixed/permeabilized cells with an excess of Purified Mouse Anti-Human TNF Cat. No. 554510; right panel prior to staining with the PE-MAb11 antibody. The quadrant markers for the bivariate dot plot was set based on the negative staining controls using isotype-matched Ig controls. Expression of TNF protein by stimulated human peripheral blood mononuclear cells PBMC . Human PBMC were stimulated for 6 hours with PMA 50 ng/ml final concentration; Sigma, Cat. #P-8139 , calcium ionophore A23187 0.5 mug/ml final concentration; Sigma, Cat. #C-9275 and PHA Gibco BRL, Cat. #10576-015 in the presence of GolgiStop 2 muM final concentration; Cat. No. 554715 . The PBMC were stained with 0.25 mug PE-Cy5 Mouse Anti-Human CD3 Cat. No. 555334 and subsequently stained with 20 mul of PE Mouse Anti-Human TNF Cat. No. 559321/554513/562083 by following the recommended assay procedure left panel . To demonstrate specificity of staining, the binding of PE-MAb11 was blocked by preincubation of the conjugate with excess 1 mug recombinant human TNF Cat. No. 554618; middle panel and by preincubation of the fixed/permeabilized cells with an excess of Purified Mouse Anti-Human TNF Cat. No. 554510; right panel prior to staining with the PE-MAb11 antibody. The quadrant markers for the bivariate dot plot was set based on the negative staining controls using isotype-matched Ig controls.