All relevant cells were harvested and rinsed utilizing FACS buffer (0.05% sodium azide and 0.5% bovine serum albumin in phosphate‐buffered saline (PBS; Thermo Fisher Scientific). For excluding dead cells, cells were firstly stained with zombie aqua reagent (#423101; Biolegend) in PBS for 10 min at 20°C in the dark. Relevant cells were stained utilizing the APC conjugated anti‐CD14 antibody (#367107; Biolegend), fluorescein isothiocyanate‐conjugated CD33 antibody (#303304; Biolegend), APC conjugated anti‐Gr‐1 antibody (#108424; Biolegend), Brilliant Violet 421 conjugated anti‐Ly6G antibody (#127628; Biolegend), Alexa Fluor® 488 conjugated anti‐CD8 antibody (#100723; Biolegend), APC‐conjugated CD4 antibody (#100412; Biolegend) in FACS buffer as mentioned for at least 30 min at 4°C in the dark. MFI of ROS was detected using CellROX™ Green Flow Cytometry Assay Kit (C10492; Thermo Fisher Scientific). Flow cytometry was applied on the LSRFortessa X‐20 (BD Biosciences). All data were analyzed utilizing FlowJo v10.1 (Treestar).
Apc conjugated cd4 antibody
Manufactured by BioLegend
Sourced in United States
The APC-conjugated CD4 antibody is a fluorescently labeled monoclonal antibody that binds to the CD4 receptor on the surface of T helper cells. It is used in flow cytometry applications to identify and quantify CD4-positive cells in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Cellrox green flow cytometry assay kit, by Thermo Fisher Scientific (1 mentions)
Zombie aqua reagent, by BioLegend (1 mentions)
Apc conjugated anti cd14 antibody, by BioLegend (1 mentions)
Lsrfortessa x 20, by BD (1 mentions)
Ficoll gradient, by GE Healthcare (1 mentions)
Cellquest software, by BD (1 mentions)
Pe conjugated anti foxp3 antibody, by BioLegend (1 mentions)
Apc conjugated anti cd11b antibody, by BioLegend (1 mentions)
Foxp3 intracellular staining kit, by BioLegend (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
2 protocols using apc conjugated cd4 antibody
1
Multiparameter Flow Cytometry Analysis
2
Quantification of T-reg and M2-monocytes in 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 (PBMCs) were isolated from 8 ml of peripheral blood (PB) using a ficoll gradient (GE healthcare, Buckinghamshire, UK). For analysis of regulatory T cells (T reg s) in PBMCs, the FoxP3 Intracellular Staining Kit (BioLegend, San Diego, CA, USA) was used; 2 x10 7 PBMCs were fixed and then incubated with PE-conjugated anti-FoxP3 antibody and APC-conjugated CD4 antibody (BioLegend, San Diego, CA, USA) for 30 min at 4 °C in the dark. To analyze the M2-skewed monocyte phenotype in PBMCs, 1 x10 7 PBMCs were incubated with an APC-conjugated anti-CD11b antibody (BioLegend) and then with a FITC-conjugated anti-CD206 antibody (Abcam) for 30 min at 4 °C, in the dark. After washing twice with FACS buffer, the proportion of T reg s and M2-phenotype monocytes in PBMCs was analyzed using a FACSCalibur flow cytometer and the CELLQuest software (Becton Dickinson, San Jose, CA, USA).
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