BD HorizonTM BV510 Rat Anti-Mouse IL-10

Description

Two-color flow cytometric analysis of IL-10 expression in stimulated mouse T cells. Splenic CD4+ T cells were cultured 2 days with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e Cat. No. 553057; 25 mug/ml for coating and soluble Purified NA/LE Hamster Anti-Mouse CD28 Cat. No. 553294; 2 mug/ml antibodies and with Recombinant IL-2 Cat. No. 550069; 10 ng/ml and IL-4 Cat. No. 550067; 25 ng/ml . The stimulated cells were subsequently cultured 3 days with Recombinant IL-2 and IL-4. The cells were then stimulated with PMA Sigma Cat. No. P-8139 and Ionomycin Sigma Cat. No. I-0634 in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . After 6 hours, the cells were harvested, washed, fixed and permeabilized with BD Cytofix/Cytoperm Fixation and Permeabilization Solution Cat. No. 554722 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with Alexa Fluor 647 Rat Anti-Mouse CD4 antibody Cat. No. 557681 and either BD Horizon BV510 Rat IgG2b, kappa Isotype Control Cat. No. 562951; Left Panel or BD Horizon BV510 Rat Anti-Mouse IL-10 antibody Cat. No. 563277; Right Panel . Flow cytometric dot plots showing correlated expression of IL-10 or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes. Flow cytometry was performed using a BD LSR II Flow Cytometer System. Two-color flow cytometric analysis of IL-10 expression in stimulated mouse T cells. Splenic CD4+ T cells were cultured 2 days with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e Cat. No. 553057; 25 mug/ml for coating and soluble Purified NA/LE Hamster Anti-Mouse CD28 Cat. No. 553294; 2 mug/ml antibodies and with Recombinant IL-2 Cat. No. 550069; 10 ng/ml and IL-4 Cat. No. 550067; 25 ng/ml . The stimulated cells were subsequently cultured 3 days with Recombinant IL-2 and IL-4. The cells were then stimulated with PMA Sigma Cat. No. P-8139 and Ionomycin Sigma Cat. No. I-0634 in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . After 6 hours, the cells were harvested, washed, fixed and permeabilized with BD Cytofix/Cytoperm Fixation and Permeabilization Solution Cat. No. 554722 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with Alexa Fluor 647 Rat Anti-Mouse CD4 antibody Cat. No. 557681 and either BD Horizon BV510 Rat IgG2b, kappa Isotype Control Cat. No. 562951; Left Panel or BD Horizon BV510 Rat Anti-Mouse IL-10 antibody Cat. No. 563277; Right Panel . Flow cytometric dot plots showing correlated expression of IL-10 or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes. Flow cytometry was performed using a BD LSR II Flow Cytometer System. Two-color flow cytometric analysis of IL-10 expression in stimulated mouse T cells. Splenic CD4+ T cells were cultured 2 days with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e Cat. No. 553057; 25 mug/ml for coating and soluble Purified NA/LE Hamster Anti-Mouse CD28 Cat. No. 553294; 2 mug/ml antibodies and with Recombinant IL-2 Cat. No. 550069; 10 ng/ml and IL-4 Cat. No. 550067; 25 ng/ml . The stimulated cells were subsequently cultured 3 days with Recombinant IL-2 and IL-4. The cells were then stimulated with PMA Sigma Cat. No. P-8139 and Ionomycin Sigma Cat. No. I-0634 in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . After 6 hours, the cells were harvested, washed, fixed and permeabilized with BD Cytofix/Cytoperm Fixation and Permeabilization Solution Cat. No. 554722 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with Alexa Fluor 647 Rat Anti-Mouse CD4 antibody Cat. No. 557681 and either BD Horizon BV510 Rat IgG2b, kappa Isotype Control Cat. No. 562951; Left Panel or BD Horizon BV510 Rat Anti-Mouse IL-10 antibody Cat. No. 563277; Right Panel . Flow cytometric dot plots showing correlated expression of IL-10 or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes. Flow cytometry was performed using a BD LSR II Flow Cytometer System. Show More Two-color flow cytometric analysis of IL-10 expression in stimulated mouse T cells. Splenic CD4+ T cells were cultured 2 days with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e Cat. No. 553057; 25 mug/ml for coating and soluble Purified NA/LE Hamster Anti-Mouse CD28 Cat. No. 553294; 2 mug/ml antibodies and with Recombinant IL-2 Cat. No. 550069; 10 ng/ml and IL-4 Cat. No. 550067; 25 ng/ml . The stimulated cells were subsequently cultured 3 days with Recombinant IL-2 and IL-4. The cells were then stimulated with PMA Sigma Cat. No. P-8139 and Ionomycin Sigma Cat. No. I-0634 in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . After 6 hours, the cells were harvested, washed, fixed and permeabilized with BD Cytofix/Cytoperm Fixation and Permeabilization Solution Cat. No. 554722 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with Alexa Fluor 647 Rat Anti-Mouse CD4 antibody Cat. No. 557681 and either BD Horizon BV510 Rat IgG2b, kappa Isotype Control Cat. No. 562951; Left Panel or BD Horizon BV510 Rat Anti-Mouse IL-10 antibody Cat. No. 563277; Right Panel . Flow cytometric dot plots showing correlated expression of IL-10 or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes. Flow cytometry was performed using a BD LSR II Flow Cytometer System. Show More Two-color flow cytometric analysis of IL-10 expression in stimulated mouse T cells. Splenic CD4+ T cells were cultured 2 days with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e Cat. No. 553057; 25 mug/ml for coating and soluble Purified NA/LE Hamster Anti-Mouse CD28 Cat. No. 553294; 2 mug/ml antibodies and with Recombinant IL-2 Cat. No. 550069; 10 ng/ml and IL-4 Cat. No. 550067; 25 ng/ml . The stimulated cells were subsequently cultured 3 days with Recombinant IL-2 and IL-4. The cells were then stimulated with PMA Sigma Cat. No. P-8139 and Ionomycin Sigma Cat. No. I-0634 in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . After 6 hours, the cells were harvested, washed, fixed and permeabilized with BD Cytofix/Cytoperm Fixation and Permeabilization Solution Cat. No. 554722 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with Alexa Fluor 647 Rat Anti-Mouse CD4 antibody Cat. No. 557681 and either BD Horizon BV510 Rat IgG2b, kappa Isotype Control Cat. No. 562951; Left Panel or BD Horizon BV510 Rat Anti-Mouse IL-10 antibody Cat. No. 563277; Right Panel . Flow cytometric dot plots showing correlated expression of IL-10 or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes. Flow cytometry was performed using a BD LSR II Flow Cytometer System. Two-color flow cytometric analysis of IL-10 expression in stimulated mouse T cells. Splenic CD4+ T cells were cultured 2 days with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e Cat. No. 553057; 25 mug/ml for coating and soluble Purified NA/LE Hamster Anti-Mouse CD28 Cat. No. 553294; 2 mug/ml antibodies and with Recombinant IL-2 Cat. No. 550069; 10 ng/ml and IL-4 Cat. No. 550067; 25 ng/ml . The stimulated cells were subsequently cultured 3 days with Recombinant IL-2 and IL-4. The cells were then stimulated with PMA Sigma Cat. No. P-8139 and Ionomycin Sigma Cat. No. I-0634 in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . After 6 hours, the cells were harvested, washed, fixed and permeabilized with BD Cytofix/Cytoperm Fixation and Permeabilization Solution Cat. No. 554722 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with Alexa Fluor 647 Rat Anti-Mouse CD4 antibody Cat. No. 557681 and either BD Horizon BV510 Rat IgG2b, kappa Isotype Control Cat. No. 562951; Left Panel or BD Horizon BV510 Rat Anti-Mouse IL-10 antibody Cat. No. 563277; Right Panel . Flow cytometric dot plots showing correlated expression of IL-10 or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes. Flow cytometry was performed using a BD LSR II Flow Cytometer System.