Cells were re-suspended in PBS containing 10% fetal bovine serum (FBS) and stained with following antibodies to analyze DC subsets in PBMC: CD3-PE-Cy™7, CD14-Alexa Fluor® 700, CD19-PE-Cy™7, CD56-PE-Cy™7, CD123-PerCP-Cy5.5, CD11c-APC and HLA-DR- APC-Cy7 (BD Bioscience). For detection of intracellular cytokine (ICC), Brefeldin A was added into culture in the last 6 hours of incubation. Cells were surface stained with subset markers and made permeable with Cytofix/cytoperm solution (BD Bioscience). Cells were then stained with following antibodies: IL-6-PE, TNF-α-Pacific Blue™, IL-1β-FITC, IL-10-PE (Biolegend) and IL-12p40/70-PE (Miltenyi Biotech). Samples were analyzed using LSRII Flow cytometer (BD Biosciences) and the data were analyzed using FlowJo software (Tree Star, Inc.)
Il 1β fitc
Manufactured by BioLegend
IL-1β-FITC is a fluorescently labeled antibody that binds to the cytokine interleukin-1 beta (IL-1β). It is designed for use in flow cytometry and other immunoassays to detect and quantify IL-1β in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Lsrii flow cytometer, by BD (1 mentions)
Cd3 pe cy7, by BD (1 mentions)
Cd11c apc, by BD (1 mentions)
Cd56 pe cy7, by BD (1 mentions)
Cd19 pe cy7, by BD (1 mentions)
Cd14 alexa fluor 700, by BD (1 mentions)
Hla dr apc cy7, by BD (1 mentions)
Cytofix cytoperm solution, by BD (1 mentions)
Tnfα pacific blue, by BioLegend (1 mentions)
Il 6 pe, by BioLegend (1 mentions)
Il 10 pe, by BioLegend (1 mentions)
Cd123 percp cy5, by BD (1 mentions)
Facscanto 2, by BD (1 mentions)
Golgiplug, by BD (1 mentions)
Cd56 pe, by BioLegend (1 mentions)
Cd16 pe cy7, by BioLegend (1 mentions)
Cd3 pe, by BioLegend (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Facs permeabilizing solution 2, by BD (1 mentions)
Il 6 apc, by BioLegend (1 mentions)
2 protocols using il 1β fitc
1
Profiling Dendritic Cell Subsets in PBMC
2
Cytokine Profile of Activated PBMCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Peripheral blood mononuclear cells were incubated overnight with 10 ng/ml LPS (Sigma-Aldrich) in the presence of GolgiPlug (1:1,000, Becton Dickinson) after pre-stimulation with IFN-γ for 2 h. The cells were then incubated with conjugated primary antibodies in phosphate-buffered saline containing 0.5% bovine serum albumine for 30 min. The antibodies used were CD3-PE, CD14-Pacfic Blue, CD16-PE-Cy7, CD20-PE, and CD56-PE (all Biolegend) at 4°C and were incubated with EDTA for 15 min followed by incubation with FACS permeabilizing solution 2 (BD Biosciences) for 15 min. Next, conjugated antibodies to TNF-α-Percp-Cy5.5, IFN-γ-APC-Cy7, IL-1β-FITC, IL-6-APC, and IL-10-APC and their respective isotype controls (all Biolegend) were added to determine intracellular cytokine production. The cells were washed and analyzed using flow cytometry (FACSCanto II, BD Biosciences) and FACSDiva software (16 (link)).
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