PBMCs (1 × 106) were Fc blocked (130-059-901; Miltenyi Biotec), stained, and acquired on a BD LSR II or a BD LSRFortessa (5 (link)). For intracellular cytokine staining, 1 × 106 PBMCs were cultured in the presence of the appropriate antiegn (ColI or ColV) in DMEM containing 5% FBS at 37°C. After overnight incubation, the cells were stimulated with PMA (10 ng/ml) and ionomycin (1 μg/ml) for 5 h, with the addition of brefeldin A (420601; BioLegend) for the last 3 h. The following Abs were used: CD3-FITC or CD3–Alexa Fluor 700 (317306/100216; BioLegend or 557943; BD Biosciences), CD11b-BV421 (562632; BD Biosciences), CD56-allophycocyanin (555518; BD Biosciences), CD14–Alexa Fluor 700 (325614; BioLegend), CD16-allophycocyanin-H7 (560195; BD Biosciences), LAIR1-PE (550811; BD Biosciences), HLA-DR–PE-Cy7 (25-9956-41; eBioscience), CD123-BV421 (306017; BioLegend), lineage-BV510 (348807; BioLegend), CD11c-allophycocyanin (559877; BD Biosciences), PE-IgG1 κ isotype control (55749; BD Biosciences), CD196/CCR6-FITC (11-1969-42; eBioscience), CD194/CCR4-PECy7 (561034; BD Biosciences), CD4-BUV395 (563552/563790; BD Biosciences), CD183/CXCR3-allophycocyanin (550967; BD Biosciences), IFN-γ-BV421 (563376; BD Biosciences), IL-17-PE (559502; BD Biosciences), γδ TCR-FITC (118105; BioLegend). Data were analyzed using FlowJo (Treestar).
Cd14 alexa fluor 700
Manufactured by BioLegend
CD14-Alexa Fluor 700 is a fluorescent-labeled antibody that specifically binds to the CD14 antigen. CD14 is a glycosylphosphatidylinositol (GPI)-anchored protein expressed on the surface of monocytes and macrophages. The Alexa Fluor 700 dye provides a bright fluorescent signal that can be detected using flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Il 17 pe, by BD (1 mentions)
Cd3 fitc, by BioLegend (1 mentions)
Brefeldin a, by BioLegend (1 mentions)
Fc blocked, by Miltenyi Biotec (1 mentions)
Cd11c allophycocyanin, by BD (1 mentions)
Cd11b bv421, by BD (1 mentions)
Cd3 alexa fluor 700, by BioLegend (1 mentions)
Cd4 buv395, by BD (1 mentions)
Ifn γ bv421, by BD (1 mentions)
Hla dr pecy7, by Thermo Fisher Scientific (1 mentions)
Cd16 apc, by BioLegend (1 mentions)
Cd11b pe cy7, by BioLegend (1 mentions)
Gm csf, by Thermo Fisher Scientific (1 mentions)
Cd34 percp cy5, by BioLegend (1 mentions)
2 protocols using cd14 alexa fluor 700
1
Multiparametric Flow Cytometry Analysis
2
Myeloid Lineage Differentiation from CD34+ Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Myeloid lineage-specific CD34+ differentiation was carried out by culturing the cells in expansion media supplemented with 20 ng/mL GM-CSF (Peprotech, ref. 300-03). Cells were cultured at 1 × 105–1 × 106 cells/mL densities and analyzed on day 7 and day 14, using CD34+-PerCP-Cy5.5 (1:60, clone 561), CD11b-PE-Cy7 (1:60, clone ICRF44), CD14-AlexaFluor700 (1:200, clone 63D3), CD16-APC (1:40, clone B73.1), and CD33-PE conjugated antibodies (1:60, clone P67.6) (all from BioLegend). Myeloid differentiation of AML cell lines was assessed using CD11b-PE-Cy7 (1:60, Biolegend clone ICRF44).
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