BD PharmingenTM APC Rat Anti-Mouse IL-6

Description

Flow cytometric analysis of intracellular IL-6 expression by activated mouse macrophages. Thioglycollate-elicited mouse peritoneal macrophages were primed with recombinant mouse IFN-gamma 10 ng/ml, Cat. No. 554587 for 2 hr and stimulated overnight with lipopolysaccharide LPS, Sigma, Cat. No. L-8272; 1 mug/ml and BD GolgiPlug Protein Transport Inhibitor Containing Brefeldin A Cat. No. 555029 . The adherent cells were washed with 1 phosphate buffered saline PBS and incubated with 1 trypsin-EDTA solution 37C, 15 min . The cells were harvested, washed, incubated with Fc Block Rat IgG2b,kappa Anti-Mouse CD16/CD32 antibody Cat. No. 553142 , fixed and permeabilized using BD Cytofix Fixation Buffer Cat. No. 554655 and BD Perm/Wash Buffer Cat. No. 554723 . The cells were then stained either with an APC Rat IgG1, kappa Isotype Control Cat No. 554686, Left Panel or with the APC Rat Anti-Mouse IL-6 antibody Cat No. 561367, Right Panel . MiCK-3 Mouse Cytokine Positive Control Cells Cat No. 554654 are prepared in a similar manner. These cells can be used as a positive control for cytokine flow cytometry experiments designed to characterize the nature of mouse IL-6-producing cells. Two-color flow cytometric dot plots showing the correlated expression of IL-6 or Ig Isotype control staining versus cellular autofluorescence measured in the phycoerythrin channel autofluorescence were derived from events with the forward and side light-scatter characteristics of intact macrophages. Flow cytometry was performed using a BD LSR II Flow Cytometer System. Flow cytometric analysis of intracellular IL-6 expression by activated mouse macrophages. Thioglycollate-elicited mouse peritoneal macrophages were primed with recombinant mouse IFN-gamma 10 ng/ml, Cat. No. 554587 for 2 hr and stimulated overnight with lipopolysaccharide LPS, Sigma, Cat. No. L-8272; 1 mug/ml and BD GolgiPlug Protein Transport Inhibitor Containing Brefeldin A Cat. No. 555029 . The adherent cells were washed with 1 phosphate buffered saline PBS and incubated with 1 trypsin-EDTA solution 37C, 15 min . The cells were harvested, washed, incubated with Fc Block Rat IgG2b,kappa Anti-Mouse CD16/CD32 antibody Cat. No. 553142 , fixed and permeabilized using BD Cytofix Fixation Buffer Cat. No. 554655 and BD Perm/Wash Buffer Cat. No. 554723 . The cells were then stained either with an APC Rat IgG1, kappa Isotype Control Cat No. 554686, Left Panel or with the APC Rat Anti-Mouse IL-6 antibody Cat No. 561367, Right Panel . MiCK-3 Mouse Cytokine Positive Control Cells Cat No. 554654 are prepared in a similar manner. These cells can be used as a positive control for cytokine flow cytometry experiments designed to characterize the nature of mouse IL-6-producing cells. Two-color flow cytometric dot plots showing the correlated expression of IL-6 or Ig Isotype control staining versus cellular autofluorescence measured in the phycoerythrin channel autofluorescence were derived from events with the forward and side light-scatter characteristics of intact macrophages. Flow cytometry was performed using a BD LSR II Flow Cytometer System. Flow cytometric analysis of intracellular IL-6 expression by activated mouse macrophages. Thioglycollate-elicited mouse peritoneal macrophages were primed with recombinant mouse IFN-gamma 10 ng/ml, Cat. No. 554587 for 2 hr and stimulated overnight with lipopolysaccharide LPS, Sigma, Cat. No. L-8272; 1 mug/ml and BD GolgiPlug Protein Transport Inhibitor Containing Brefeldin A Cat. No. 555029 . The adherent cells were washed with 1 phosphate buffered saline PBS and incubated with 1 trypsin-EDTA solution 37C, 15 min . The cells were harvested, washed, incubated with Fc Block Rat IgG2b,kappa Anti-Mouse CD16/CD32 antibody Cat. No. 553142 , fixed and permeabilized using BD Cytofix Fixation Buffer Cat. No. 554655 and BD Perm/Wash Buffer Cat. No. 554723 . The cells were then stained either with an APC Rat IgG1, kappa Isotype Control Cat No. 554686, Left Panel or with the APC Rat Anti-Mouse IL-6 antibody Cat No. 561367, Right Panel . MiCK-3 Mouse Cytokine Positive Control Cells Cat No. 554654 are prepared in a similar manner. These cells can be used as a positive control for cytokine flow cytometry experiments designed to characterize the nature of mouse IL-6-producing cells. Two-color flow cytometric dot plots showing the correlated expression of IL-6 or Ig Isotype control staining versus cellular autofluorescence measured in the phycoerythrin channel autofluorescence were derived from events with the forward and side light-scatter characteristics of intact macrophages. Flow cytometry was performed using a BD LSR II Flow Cytometer System. Show More Flow cytometric analysis of intracellular IL-6 expression by activated mouse macrophages. Thioglycollate-elicited mouse peritoneal macrophages were primed with recombinant mouse IFN-gamma 10 ng/ml, Cat. No. 554587 for 2 hr and stimulated overnight with lipopolysaccharide LPS, Sigma, Cat. No. L-8272; 1 mug/ml and BD GolgiPlug Protein Transport Inhibitor Containing Brefeldin A Cat. No. 555029 . The adherent cells were washed with 1 phosphate buffered saline PBS and incubated with 1 trypsin-EDTA solution 37C, 15 min . The cells were harvested, washed, incubated with Fc Block Rat IgG2b,kappa Anti-Mouse CD16/CD32 antibody Cat. No. 553142 , fixed and permeabilized using BD Cytofix Fixation Buffer Cat. No. 554655 and BD Perm/Wash Buffer Cat. No. 554723 . The cells were then stained either with an APC Rat IgG1, kappa Isotype Control Cat No. 554686, Left Panel or with the APC Rat Anti-Mouse IL-6 antibody Cat No. 561367, Right Panel . MiCK-3 Mouse Cytokine Positive Control Cells Cat No. 554654 are prepared in a similar manner. These cells can be used as a positive control for cytokine flow cytometry experiments designed to characterize the nature of mouse IL-6-producing cells. Two-color flow cytometric dot plots showing the correlated expression of IL-6 or Ig Isotype control staining versus cellular autofluorescence measured in the phycoerythrin channel autofluorescence were derived from events with the forward and side light-scatter characteristics of intact macrophages. Flow cytometry was performed using a BD LSR II Flow Cytometer System. Show More Flow cytometric analysis of intracellular IL-6 expression by activated mouse macrophages. Thioglycollate-elicited mouse peritoneal macrophages were primed with recombinant mouse IFN-gamma 10 ng/ml, Cat. No. 554587 for 2 hr and stimulated overnight with lipopolysaccharide LPS, Sigma, Cat. No. L-8272; 1 mug/ml and BD GolgiPlug Protein Transport Inhibitor Containing Brefeldin A Cat. No. 555029 . The adherent cells were washed with 1 phosphate buffered saline PBS and incubated with 1 trypsin-EDTA solution 37C, 15 min . The cells were harvested, washed, incubated with Fc Block Rat IgG2b,kappa Anti-Mouse CD16/CD32 antibody Cat. No. 553142 , fixed and permeabilized using BD Cytofix Fixation Buffer Cat. No. 554655 and BD Perm/Wash Buffer Cat. No. 554723 . The cells were then stained either with an APC Rat IgG1, kappa Isotype Control Cat No. 554686, Left Panel or with the APC Rat Anti-Mouse IL-6 antibody Cat No. 561367, Right Panel . MiCK-3 Mouse Cytokine Positive Control Cells Cat No. 554654 are prepared in a similar manner. These cells can be used as a positive control for cytokine flow cytometry experiments designed to characterize the nature of mouse IL-6-producing cells. Two-color flow cytometric dot plots showing the correlated expression of IL-6 or Ig Isotype control staining versus cellular autofluorescence measured in the phycoerythrin channel autofluorescence were derived from events with the forward and side light-scatter characteristics of intact macrophages. Flow cytometry was performed using a BD LSR II Flow Cytometer System. Flow cytometric analysis of intracellular IL-6 expression by activated mouse macrophages. Thioglycollate-elicited mouse peritoneal macrophages were primed with recombinant mouse IFN-gamma 10 ng/ml, Cat. No. 554587 for 2 hr and stimulated overnight with lipopolysaccharide LPS, Sigma, Cat. No. L-8272; 1 mug/ml and BD GolgiPlug Protein Transport Inhibitor Containing Brefeldin A Cat. No. 555029 . The adherent cells were washed with 1 phosphate buffered saline PBS and incubated with 1 trypsin-EDTA solution 37C, 15 min . The cells were harvested, washed, incubated with Fc Block Rat IgG2b,kappa Anti-Mouse CD16/CD32 antibody Cat. No. 553142 , fixed and permeabilized using BD Cytofix Fixation Buffer Cat. No. 554655 and BD Perm/Wash Buffer Cat. No. 554723 . The cells were then stained either with an APC Rat IgG1, kappa Isotype Control Cat No. 554686, Left Panel or with the APC Rat Anti-Mouse IL-6 antibody Cat No. 561367, Right Panel . MiCK-3 Mouse Cytokine Positive Control Cells Cat No. 554654 are prepared in a similar manner. These cells can be used as a positive control for cytokine flow cytometry experiments designed to characterize the nature of mouse IL-6-producing cells. Two-color flow cytometric dot plots showing the correlated expression of IL-6 or Ig Isotype control staining versus cellular autofluorescence measured in the phycoerythrin channel autofluorescence were derived from events with the forward and side light-scatter characteristics of intact macrophages. Flow cytometry was performed using a BD LSR II Flow Cytometer System.