The total ROS production was detected by H2DCFDA, according to the manufacturer's instructions (Invitrogen). Cells were quantified by flow cytometry following manufacturer's instructions. Data were analysed by FlowJo (Tree Star, Inc.). For detection of neutrophils or intracellular TNF‐α, cells were stained with APC‐labelled rat anti‐mouse CD45 antibody (1:100; BD Biosciences, Franklin Lakes, NJ, USA), PerCP‐Cy5.5‐labelled rat anti‐mouse CD11b antibody (1:100; BD Biosciences) and FITC‐labelled rat anti‐mouse Ly6G antibody (1:100; BD Biosciences) for 30 minutes in PBS at 4°C and then washed twice with PBS. Then, the cells were fixed in 4% paraformaldehyde solution for 20 minutes, permeabilized with 0.5% Tritox‐100 for 30 minutes at 4°C and washed with PBS. Next, the cells were stained with a PE‐labelled rat anti‐mouse TNF‐α antibody (1:100; BD Biosciences) for 2 hours at 4°C and then washed. Data acquisition was performed on an Aria III flow cytometer, and the results were analysed by Flowjo, Version 7.6.1; Treestar, Ashland, OR, USA.
Pe labelled rat anti mouse tnf α antibody
Manufactured by BD
The PE-labelled rat anti-mouse TNF-α antibody is a laboratory reagent used for the detection and quantification of mouse tumor necrosis factor-alpha (TNF-α) in biological samples. This antibody is conjugated with the fluorescent dye phycoerythrin (PE), which allows for the visualization and measurement of TNF-α levels through flow cytometry or other fluorescence-based techniques.
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Lab products found in correlation
Apc labelled rat anti mouse cd45 antibody, by BD (1 mentions)
Percp cy5.5 labelled rat anti mouse cd11b antibody, by BD (1 mentions)
Aria 3 flow cytometer, by Tree Star (1 mentions)
H2dcfda, by Thermo Fisher Scientific (1 mentions)
Apc labelled rat anti mouse cd45, by BD (1 mentions)
Percp cy5.5 labelled rat anti mouse cd11b, by BD (1 mentions)
2 protocols using pe labelled rat anti mouse tnf α antibody
1
Quantifying Cellular ROS and Inflammatory Markers
2
Neutrophil-Monocyte Co-culture Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Neutrophils (1×106 cells/well) were co-cultured with monocytes (5×105 cells/well) and stimulated with necrotic lung cells (1×105 cells/mL) or mitochondrial DNA (5 μg/mL) in a 24-well plate at 37 °C for 4 h. Neutrophils were stained with APC-labelled rat anti-mouse CD45 (BD Biosciences, 1:100), PerCP-Cy5.5-labelled rat anti-mouse CD11b (BD Biosciences, 1:100) and FITC-labelled rat anti-mouse Ly6G antibodies (BD Biosciences, 1:100) for 30 min in PBS at 4 °C and then were washed twice with PBS. Cells were fixed in 2% paraformaldehyde solution for 20 min, permeabilized using 1% Triton X-100 for 30 min at 4 °C and washed with PBS. The cells were stained with PE-labelled rat anti-mouse TNF-α antibody (BD Biosciences, 1:100) for 2 h at 4 °C and then were washed. Data acquisition was performed on a FACS AriaIII flow cytometer, and the results were analysed by FlowJo software.
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