The phenotype of cells from mouse lungs was determined by incubating lung leukocytes with the indicated antibodies and CD16/32 to limit nonspecific binding. Leukocytes were stained at 4°C for 15 min in PBS containing 1%BSA and 0.01% sodium azide. Cells were stained with combinations of the following antibodies: FITC-conjugated I-Ab; PE-conjugated CD11c; PerCP-conjugated CD45; and APC-conjugated F4/80 and EpCAM from BD Biosciences. For intracellular IL-33 staining, cells were incubated with Cytofix/Cytoperm (BD Biosciences, San Diego, CA), washed in Permeabilization Buffer (BD Biosciences), and stained for 30 min with PE-conjugated IL-33 (R&D systems, Minneapolis, MN). Cells were washed and resuspended in 1% paraformaldehyde. Isotype controls were used. Data was acquired using BD Accuri™ C6 cytometer and analyzed using FCS Express 4.0 Software. For cell sorting experiments, F4/80+ leukocytes from the lungs of WT and CCR2−/− mice were isolated at day-7 p.i. using 5-laser FACS Aria II (BD Biosciences).
5 laser facs aria 2
Manufactured by BD
The BD 5-laser FACS Aria II is a high-performance cell sorter that utilizes five lasers to enable the detection and sorting of multiple cellular parameters simultaneously. The core function of this instrument is to provide researchers with the ability to efficiently analyze and isolate specific cell populations from complex samples.
Automatically generated - may contain errors
Lab products found in correlation
Permeabilization buffer, by BD (2 mentions)
Fitc conjugated 1 ab, by BD (2 mentions)
Pe conjugated cd11c, by BD (2 mentions)
Apc conjugated f4 80 and epcam, by BD (2 mentions)
Pe conjugated il 33, by R&D Systems (2 mentions)
Percp conjugated cd45, by BD (2 mentions)
Accuri c6 cytometer, by BD (2 mentions)
Cytofix cytoperm, by BD (2 mentions)
2 protocols using 5 laser facs aria 2
1
Lung Leukocyte Phenotyping in Mice
2
Lung Leukocyte Phenotyping in Mice
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The phenotype of cells from mouse lungs was determined by incubating lung leukocytes with the indicated antibodies and CD16/32 to limit nonspecific binding. Leukocytes were stained at 4°C for 15 min in PBS containing 1%BSA and 0.01% sodium azide. Cells were stained with combinations of the following antibodies: FITC-conjugated I-Ab; PE-conjugated CD11c; PerCP-conjugated CD45; and APC-conjugated F4/80 and EpCAM from BD Biosciences. For intracellular IL-33 staining, cells were incubated with Cytofix/Cytoperm (BD Biosciences, San Diego, CA), washed in Permeabilization Buffer (BD Biosciences), and stained for 30 min with PE-conjugated IL-33 (R&D systems, Minneapolis, MN). Cells were washed and resuspended in 1% paraformaldehyde. Isotype controls were used. Data was acquired using BD Accuri™ C6 cytometer and analyzed using FCS Express 4.0 Software. For cell sorting experiments, F4/80+ leukocytes from the lungs of WT and CCR2−/− mice were isolated at day-7 p.i. using 5-laser FACS Aria II (BD Biosciences).
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