Peripheral blood mononuclear cells (PBMC) were isolated from the peripheral blood samples of SLE patients by using Lymphoprep™ (STEMCELL). Freshly isolated PBMCs were stained for surface marker FITC anti-human Lineage Cocktail (CD3, CD14, CD19, CD20, CD56) (BD Pharmingen), PerCp anti-human HLA-DR (BD Pharmingen), PE anti-human CD11c (BD Pharmingen), APC anti-human CD40 (BD Pharmingen) or APC anti-human CD86 (BD Pharmingen). After surface staining, cells were fixed/permeabilized with fixation/permeabilization solution (Cytofix/Cytoperm™; BD Pharmingen), and stained with p-mTOR-Alexa 647 (BD Pharmingen) for 30 min at 4°C. Cells were then washed with 1 × Perm/Wash Buffer (BD Pharmingen) and resuspended in PBS + 2% FBS for flow cytometric analysis. Flow cytometry was performed on a BD FACS Canto II (BD Biosciences) and data were analyzed using FCS Express 4 software (De Novo Software, Los Angeles, CA).
Apc anti human cd86
Manufactured by BD
APC anti-human CD86 is a fluorescently labeled antibody that binds to the CD86 protein expressed on the surface of various cell types. CD86 is a co-stimulatory molecule involved in the activation of T cells. The APC fluorescent label allows for the detection and analysis of CD86-positive cells using flow cytometry or other fluorescence-based techniques.
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Lab products found in correlation
2 protocols using apc anti human cd86
1
Analyzing Immune Cell Activation in SLE
2
Inflammatory Response Assay in THP-1 and PBMC
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THP-1 cells or PBMC (1 ×
105 cells/well) were plated
in 200 μL/well in 96-well round-bottom plates and treated with
compounds overnight or compound plus LPS (10 ng/mL) or MPLA (1 μg/mL).
For treatment with inhibitors, cells were pre-incubated with each
inhibitor for 1 h prior to exposure to the compounds. The concentrations
of compounds and inhibitors are detailed in each figure legend.
CXCL8, CCL-3, and TNF-α in the supernatant were tested by ELISA.
CD40 and CD86 expression was measured using PE anti-human CD40 (BioLegends,
#313006) and APC anti-human CD86 (BD bioscience #560956) by MAQSQuant
Analyzer 20 (Miltenyi Biotec) and FlowJo (version 10.6.1, Becton Dickinson).
105 cells/well) were plated
in 200 μL/well in 96-well round-bottom plates and treated with
compounds overnight or compound plus LPS (10 ng/mL) or MPLA (1 μg/mL).
For treatment with inhibitors, cells were pre-incubated with each
inhibitor for 1 h prior to exposure to the compounds. The concentrations
of compounds and inhibitors are detailed in each figure legend.
CXCL8, CCL-3, and TNF-α in the supernatant were tested by ELISA.
CD40 and CD86 expression was measured using PE anti-human CD40 (BioLegends,
#313006) and APC anti-human CD86 (BD bioscience #560956) by MAQSQuant
Analyzer 20 (Miltenyi Biotec) and FlowJo (version 10.6.1, Becton Dickinson).
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