Apc conjugated hladr

Manufactured by BioLegend

APC-conjugated HLADR is a fluorescent-labeled antibody that binds to the HLADR protein on the surface of human cells. It can be used in flow cytometry applications to identify and quantify HLADR-expressing cells.

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Lab products found in correlation

Fitc conjugated cd86, by BD (1 mentions) Cd14 magnetic beads, by Miltenyi Biotec (1 mentions) Pe conjugated cd14, by BioLegend (1 mentions) Canto 2 flow cytometer, by BD (1 mentions) Apc cy7 conjugated anti cd14, by BioLegend (1 mentions) Pacific blue conjugated anti cd3, by BioLegend (1 mentions) Pe cy7 conjugated anti cd19, by BioLegend (1 mentions)

2 protocols using apc conjugated hladr

Monocyte Phenotypic Characterization

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Human monocytes isolated with CD14 magnetic beads (Miltenyi Biotec) from human PBMCs were exposed for 5 days in the presence of ACM. Cells were then collected and stained with APC-conjugated CCR2 (Biolegend, diluted 1:50), PE-conjugated CD14 (Biolegend, diluted 1:100), APC-conjugated HLADR (Biolegend, diluted 1:50), FITC-conjugated CD80 (BD pharmingen, diluted 1:50) and FITC-conjugated CD86 (BD pharmingen, diluted 1:50) after the FcR block for 15 min (Miltenyl Biotec, diluted 1:100). In high CD14 expressing cells, we examined the intensity with the other antibody. The mean fluorescence intensity (MFI) of each sample was analyzed with Canto II flow cytometer (BD biosciences).

Shimizu M., Okuno T., Kinoshita M., Sumi H., Fujimura H., Yamashita K., Sugimoto T., Sakakibara S., Sakakibara K., Koda T., Tada S., Ishikura T., Murata H., Beppu S., Shiraishi N., Sugiyama Y., Nakatsuji Y., Kumanogoh A, & Mochizuki H. (2020). Mitochondrial DNA enhance innate immune responses in neuromyelitis optica by monocyte recruitment and activation. Scientific Reports, 10, 13274.

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Multiparametric Flow Cytometry Profiling of PBMCs

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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.

Chowdhury R.R., Vallania F., Yang Q., Angel C.J., Darboe F., Penn-Nicholson A., Rozot V., Nemes E., Malherbe S.T., Ronacher K., Walzl G., Hanekom W., Davis M.M., Winter J., Chen X., Scriba T.J., Khatri P, & Chien Y.H. (2018). A multi-cohort study of the immune factors associated with M. tuberculosis infection outcomes. Nature, 560(7720), 644-648.

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