Cells were labeled using the anti-mouse antibodies: CD11b-PE (concentration 1:400), iNOS-Alexa Fluor 488 (concentration 1:100), and CD206-APC (concentration 1:100) (Biolegend). We collected 10,000 cell events per sample, and we performed unstained control as well as isotype controls. Labeled cells were examined on the LSR II Analyzer (BD) in the Stanford Shared FACS Facility, and data was analyzed with FlowJo software (Tree Star, Ashland, OR). Dead cells were excluded by ethidium monoazide (EMA) staining, and appropriate isotype controls were used. Quadrants were drawn using the M0 isotope control.
Inos alexa fluor 488
Manufactured by BioLegend
The INOS-Alexa Fluor 488 is a fluorescently labeled antibody that specifically recognizes the inducible nitric oxide synthase (iNOS) protein. Alexa Fluor 488 is the fluorescent dye conjugated to the antibody, which emits green fluorescence upon excitation. This product can be used in flow cytometry, immunofluorescence, and other applications that require the detection of iNOS expression.
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Lab products found in correlation
2 protocols using inos alexa fluor 488
1
Multicolor Flow Cytometry Analysis
2
Macrophage Phenotyping by Flow Cytometry
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Macrophage phenotypes in the monoculture and cocultures were analyzed by flow cytometry 72 hours after seeding or the last IL-4 treatment, whichever was later. Cells were suspended in 2 % FBS/PBS, preincubated with anti-CD16/32 mAb to prevent nonspecific binding via FcRII/III interactions, and incubated with anti-mouse mAb (CD11b-PE, iNOS-Alexa Fluor 488, and CD206-APC; Biolegend). Appropriate isotypes were used and ethidium monoazide bromide staining excluded dead cells. Analysis was performed on the LSR II Analyzer (BD Immunocytometry Systems, San Diego, CA, USA) in the Stanford Shared FACS Facility, using FlowJo software (TreeStar, Ashland, OR, USA).
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