For flow cytometry experiments, PBMCs were thawed and rested as described in ‘Mass cytometry measurements and analysis’. Cells (3–5 × 106) were incubated with antibodies for 30 min at 4 °C, then washed with FACS staining buffer (PBS containing 1% bovine serum albumin and 0.05% sodium azide). The following monoclonal antibodies were used: Pacific Blue-conjugated anti-CD3 (BioLegend, 300417), PE–Cy7-conjugated anti-CD19 (BioLegend, 302216), APC–Cy7-conjugated anti-CD14 (BioLegend, 325620), APC-conjugated HLA-DR (BioLegend, 307610), PE–Dazzle-conjugated anti-CD16 (BioLegend, 302054) and Brilliant Violet 785-conjugated CD56 (BioLegend, 362550). The live/dead aqua-amine reactive dye was used for gating dead cells. All antibodies were validated by the manufacturers for flow cytometry application, as indicated on the manufacturer’s website. Data were analysed using FlowJo version 10.2.
Apc cy7 conjugated anti cd14
Manufactured by BioLegend
APC–Cy7-conjugated anti-CD14 is a fluorochrome-conjugated antibody that recognizes the CD14 protein. CD14 is a surface glycoprotein that serves as a co-receptor for the detection of bacterial lipopolysaccharide.
Automatically generated - may contain errors
Lab products found in correlation
Apc conjugated hladr, by BioLegend (1 mentions)
Pacific blue conjugated anti cd3, by BioLegend (1 mentions)
Pe cy7 conjugated anti cd19, by BioLegend (1 mentions)
Pe conjugated anti ifn γ, by BioLegend (1 mentions)
Peptivator, by Miltenyi Biotec (1 mentions)
Apc cy7 conjugated anti cd19, by BioLegend (1 mentions)
Pe cy7 conjugated anti cd8, by BioLegend (1 mentions)
Lsrfortessa, by BD (1 mentions)
Cytofix cytoperm, by BD (1 mentions)
Golgistop, by BD (1 mentions)
Golgiplug, by BD (1 mentions)
2 protocols using apc cy7 conjugated anti cd14
1
Multiparametric Flow Cytometry Profiling of PBMCs
2
SARS-CoV-2 Spike Protein T-Cell Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Overnight-rested PBMCs were stimulated with SARS-CoV-2 overlapping peptide pools against SARS-CoV-2 spike protein (PepTivator, Miltenyi Biotec) at a final concentration of 1 μg ml -1 for 1 h in the presence of 1 μg ml -1 monoclonal antibodies CD28 and CD49d, and then for an additional 5 h with GolgiPlug and GolgiStop (BD Biosciences). Dead cells were labelled using Fixable Viability eFluor 780 dye (Thermo Fisher Scientific). Surface markers were stained using BV786-conjugated anti-CD3 (BD Biosciences, 565491; diluted 1:100), BUV486-conjugated anti-CD4 (BD Biosciences, 612937; 1:50), PE-Cy7-conjugated anti-CD8 (BioLegend, 301012; 1:100), APC-Cy7-conjugated anti-CD14 (BioLegend, 301820; 1:100), APC-Cy7-conjugated anti-CD56 (BioLegend, 362512; 1:100) and APC-Cy7-conjugated anti-CD19 (BioLegend, 302218; 1:100). Cells were then washed, fixed with Cytofix/Cytoperm (BD Biosciences) and stained with PE-conjugated anti-IFNγ (BioLegend). Negative controls without peptide stimulation were run for each sample. All results were acquired on a BD LSRFortessa (BD Biosciences) flow cytometer using the BD FACSDIVA v.8.01 software and analysed using FlowJo v.10.6.1 software.
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