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.
Percp cy5.5 labelled rat anti mouse cd11b antibody
Manufactured by BD
Sourced in United States
PerCP‐Cy5.5‐labelled rat anti‐mouse CD11b antibody is a laboratory reagent used for the detection and identification of CD11b-expressing cells in flow cytometry analysis. The antibody is conjugated with the PerCP‐Cy5.5 fluorescent dye, which allows for the simultaneous detection of CD11b-positive cells alongside other markers.
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Lab products found in correlation
Apc labelled rat anti mouse cd45 antibody, by BD (2 mentions)
Pe labelled rat anti mouse tnf α antibody, by BD (1 mentions)
Aria 3 flow cytometer, by Tree Star (1 mentions)
H2dcfda, by Thermo Fisher Scientific (1 mentions)
Cpg1668, by Thermo Fisher Scientific (1 mentions)
Golgiplug, by BD (1 mentions)
2 protocols using percp cy5.5 labelled rat anti mouse cd11b antibody
1
Quantifying Cellular ROS and Inflammatory Markers
2
Cytokine Profiling of Monocytes Stimulated by Necrotic Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Bone marrow monocytes (1×106 cells/well) were stimulated with necrotic lung cells (1×105 cells/mL), mitochondrial DNA (5 μg/mL) or CpG1668 (1 μg/mL) (Invitrogen) in the presence of brefeldin A (GolgiPlug, BD Biosciences) in a 24-well plate at 37 °C for 4 h. For intracellular cytokine detection, the cells were stained with APC-labelled rat anti-mouse CD45 antibody (BD Biosciences, 1:100), PerCP-Cy5.5-labelled rat anti-mouse CD11b antibody (BD Biosciences, 1:100) and PE-labelled rat anti-mouse Ly6C antibody (BD Biosciences, 1:100) for 30 min at 4 °C and then were fixed and permeabilized as described. The intracellular cytokines were stained with FITC-labelled rat anti-mouse TNF-α antibody (BD Biosciences, 1:100), Brilliant Violet 421TM rat anti-mouse IL-10 antibody (BD Biosciences, 1:100), FITC-labelled rat anti-mouse IL-6 antibody (BD Biosciences, 1:100) and FITC-labelled rat anti-mouse IFN-γ antibody (BD Biosciences, 1:100) or their appropriate isotype controls, including rat IgG1 and IgG2b, for 2 h in PBS at 4 °C. Data acquisition was performed on a FACS AriaIII flow cytometer, and the results were analysed by FlowJo software.
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