BD PharmingenTM Alexa Fluor 488 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 hrs 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 PE-conjugated rat anti-mouse CD4 PE-RM4-5, Cat. No. 553048 and either rat anti-mouse TNF antibody Alexa Fluor 488-MP6-XT22, Cat. No. 557719 , left panel or immunoglobulin isotype control Alexa 488-R3-34, Cat. No. 557720 , right panel by using Pharmingens staining protocol. To demonstrate specificity of staining the binding of Alexa Fluor 488-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 MP6XT22 antibody 5 mug, Cat. No. 554416, data not shown prior to stainining. Dot plots were derived from gated events with the forward and side light scatter characteristics of lymphocytes. The quadarant 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 hrs 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 PE-conjugated rat anti-mouse CD4 PE-RM4-5, Cat. No. 553048 and either rat anti-mouse TNF antibody Alexa Fluor 488-MP6-XT22, Cat. No. 557719 , left panel or immunoglobulin isotype control Alexa 488-R3-34, Cat. No. 557720 , right panel by using Pharmingens staining protocol. To demonstrate specificity of staining the binding of Alexa Fluor 488-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 MP6XT22 antibody 5 mug, Cat. No. 554416, data not shown prior to stainining. Dot plots were derived from gated events with the forward and side light scatter characteristics of lymphocytes. The quadarant 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 hrs 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 PE-conjugated rat anti-mouse CD4 PE-RM4-5, Cat. No. 553048 and either rat anti-mouse TNF antibody Alexa Fluor 488-MP6-XT22, Cat. No. 557719 , left panel or immunoglobulin isotype control Alexa 488-R3-34, Cat. No. 557720 , right panel by using Pharmingens staining protocol. To demonstrate specificity of staining the binding of Alexa Fluor 488-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 MP6XT22 antibody 5 mug, Cat. No. 554416, data not shown prior to stainining. Dot plots were derived from gated events with the forward and side light scatter characteristics of lymphocytes. The quadarant 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 hrs 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 PE-conjugated rat anti-mouse CD4 PE-RM4-5, Cat. No. 553048 and either rat anti-mouse TNF antibody Alexa Fluor 488-MP6-XT22, Cat. No. 557719 , left panel or immunoglobulin isotype control Alexa 488-R3-34, Cat. No. 557720 , right panel by using Pharmingens staining protocol. To demonstrate specificity of staining the binding of Alexa Fluor 488-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 MP6XT22 antibody 5 mug, Cat. No. 554416, data not shown prior to stainining. Dot plots were derived from gated events with the forward and side light scatter characteristics of lymphocytes. The quadarant 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 hrs 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 PE-conjugated rat anti-mouse CD4 PE-RM4-5, Cat. No. 553048 and either rat anti-mouse TNF antibody Alexa Fluor 488-MP6-XT22, Cat. No. 557719 , left panel or immunoglobulin isotype control Alexa 488-R3-34, Cat. No. 557720 , right panel by using Pharmingens staining protocol. To demonstrate specificity of staining the binding of Alexa Fluor 488-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 MP6XT22 antibody 5 mug, Cat. No. 554416, data not shown prior to stainining. Dot plots were derived from gated events with the forward and side light scatter characteristics of lymphocytes. The quadarant 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 hrs 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 PE-conjugated rat anti-mouse CD4 PE-RM4-5, Cat. No. 553048 and either rat anti-mouse TNF antibody Alexa Fluor 488-MP6-XT22, Cat. No. 557719 , left panel or immunoglobulin isotype control Alexa 488-R3-34, Cat. No. 557720 , right panel by using Pharmingens staining protocol. To demonstrate specificity of staining the binding of Alexa Fluor 488-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 MP6XT22 antibody 5 mug, Cat. No. 554416, data not shown prior to stainining. Dot plots were derived from gated events with the forward and side light scatter characteristics of lymphocytes. The quadarant markers for the bivariate dot plots were set based on the autofluorescence and isotype controls.