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Hla dr vioblue

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HLA-DR-VioBlue is a monoclonal antibody conjugated with the fluorescent dye VioBlue. It is used for the detection and quantification of HLA-DR expression on the surface of cells by flow cytometry.

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Lab products found in correlation

Macsquantify software, by Miltenyi Biotec (3 mentions) Pan flavivirus 4g2 antibody, by Merck Group (2 mentions) Goat anti mouse 488, by BD (2 mentions) Dc sign apc, by BD (2 mentions) Macsquant, by Miltenyi Biotec (2 mentions) Cd38 pevio770, by Miltenyi Biotec (1 mentions) Granzyme b pe, by Miltenyi Biotec (1 mentions) Perforin fitc, by Miltenyi Biotec (1 mentions) Facsverse cytometer, by BD (1 mentions) Cd4 apc vio770, by Miltenyi Biotec (1 mentions) Cd8 apc, by Miltenyi Biotec (1 mentions) Lsr fortessa cytometer, by BD (1 mentions) Cd56 bv510, by BioLegend (1 mentions) Anti cd14 bv650, by BioLegend (1 mentions) Cd19 viogreen, by Miltenyi Biotec (1 mentions) Cd16 pe, by Beckman Coulter (1 mentions) Viobility 405 520, by Miltenyi Biotec (1 mentions) Anti cd3 bv510, by BD (1 mentions) Anti cd73 pe, by Miltenyi Biotec (1 mentions) Anti cd133 apc, by Miltenyi Biotec (1 mentions)

6 protocols using hla dr vioblue

1

Flavivirus Infection Analysis in Cells

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The HC, trophoblasts, or U251 cells were infected as described above. Cells were fixed using CytoFix/Cytoperm and permeabilized according to the manufacturer's protocol (BD Biosciences, San Jose, CA), followed by staining with pan flavivirus 4G2 antibody (Millipore, Billerica, MA) and secondary staining with goat anti-mouse 488 (BD Biosciences). The HC were also stained with HLA-DR-VioBlue (Miltenyi Biotec) and DC-SIGN-APC (BD Biosciences), and 4G2+ cells, together with HLA-DR+ and DC-SIGN+ cells were quantified with flow cytometry (MacsQuant, Miltenyi Biotec). FACS plots were established based on forward and side scatter, doublet discrimination, and gates were set based on uninfected, stained control cell samples. Analysis was performed with MacsQuantify software (Miltenyi Biotec).
Gavegnano C., Bassit L.C., Cox B.D., Hsiao H.M., Johnson E.L., Suthar M., Chakraborty R, & Schinazi R.F. (2017). Jak Inhibitors Modulate Production of Replication-Competent Zika Virus in Human Hofbauer, Trophoblasts, and Neuroblastoma cells. Pathogens & Immunity, 2(2), 199-218.
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2

Phenotyping of Activated PBMC Subsets

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1.5 × 10^6 of thawed PBMCs were plated in complete RPMI containing 10% human serum supplemented with 1% Penicillin–Streptomycin–Glutamin. Overnight-rested PBMCs were stained with the appropriate antibodies for 20 min at 4 °C in the dark and acquired using FACSVerse™cytometer (BD Biosciences). Dead cells were labeled using ViobilityTM Fixable Dye (Miltenyi Biotec). Antibodies used were: CD4-APC-Vio770, CD8-APC, HLA-DR-VioBlue, CD38-PE-Vio770, Granzyme B-PE and Perforin-FITC (Miltenyi Biotec). (Representative plots are shown in Supplementary Fig. 6). Data were analyzed using FlowJo 10.7.2 (BD Biosciences).
Rovito R., Bono V., Augello M., Tincati C., Mainoldi F., Beaudoin-Bussières G., Tauzin A., Bianchi S., Hadla M., Yellenki V., d’Arminio Monforte A., Casola S., Borghi E., Finzi A, & Marchetti G. (2022). Association between SARS-CoV-2 RNAemia and dysregulated immune response in acutely ill hospitalized COVID-19 patients. Scientific Reports, 12, 19658.
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3

Phenotypic analysis of dendritic cells

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Frozen PBMC were thawed in RPMI-50% FCS, counted in Trypan blue, and 1 × 106 cells were stained with a viability dye (Viobility 405/520, Fixable Dye, Miltenyi-Biotech), Lineage antibodies (anti-CD3-BV510 (BD Biosciences), CD19-VioGreen (Miltenyi-Biotech) and CD56-BV510 (BioLegend)), and anti-CD14-BV650 (Biolegend), HLA-DR VioBlue (Miltenyi-Biotech), CD141-BV785 (BioLegend), CD16-PE (Beckman-Coulter), CD1c-BV605 (BD Biosciences), FcERI-PerCP-Vio700 (Miltenyi-Biotech), CD123-APC-Vio770 (Miltenyi-Biotech), CD11c-PE-Vio615 (Miltenyi-Biotech) antibodies (all antibodies are described in Additional file 1: Table S3). Samples were processed on the BD LSRFortessa cytometer (BD Biosciences). pDC and DC cells were identified as described by Mair et al. [49 (link)]. Data were analyzed using FlowJoTM version v10·8 software (BD Life Sciences).
Garcia G., Labrouche-Colomer S., Duvignaud A., Clequin E., Dussiau C., Trégouët D.A., Malvy D., Prevel R., Zouine A., Pellegrin I., Goret J., Mamani-Matsuda M., Dewitte A, & James C. (2024). Impaired balance between neutrophil extracellular trap formation and degradation by DNases in COVID-19 disease. Journal of Translational Medicine, 22, 246.
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4

Immunophenotyping of Mesenchymal Progenitor Cells

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Freshly isolated MPCs and P2-MSCs were washed in MACSQuant™ Running Buffer (Miltenyi Biotech, Bergisch Gladbach, Germany) and stained with anti-CD11c VioBlue®, anti-CD18 APC, anti-CD31 PE, anti-CD34 VioBlue®, anti-CD45 APC-Vio770, anti-CD73 PE, anti-CD90 FITC, anti-CD133 APC, anti-CD146 FITC, HLA-DR VioBlue® (Miltenyi Biotech), anti-STRO-1 FITC, and CD144 PE (Biolegend, San Diego, CA, USA). Samples were acquired by MACSQuant® Flow Cytometer and analyzed by MACSQuantify® Software (Miltenyi Biotech).
Montali M., Panvini F.M., Barachini S., Ronca F., Carnicelli V., Mazzoni S., Petrini I, & Pacini S. (2017). Human adult mesangiogenic progenitor cells reveal an early angiogenic potential, which is lost after mesengenic differentiation. Stem Cell Research & Therapy, 8, 106.
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5

Characterization of ADSVCS Surface Markers

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Freshly isolated cells were characterized for ADSVCs surface protein expression [17 (link)] by flow cytometry (MACSQuant analyzer, Miltenyi-Biotech) according to the manufacturer’s instructions. Cells were stained with the following antihuman-conjugated monoclonal antibodies: CD13-APC, CD14-PE, CD29-FITC, CD31-APC, CD34-PE, CD45-VioBlue, CD73-APC, CD90-FITC, CD105-VioBlue, CD144 (VE-Cadherin)-PE, CD146-Biotin, CD166-Biotin, HLA-ABC-FITC, HLA-DR-VioBlue, or relevant isotype-matched controls (Miltenyi-Biotech). Isotypes controls and automated compensation were settled to minimize false positive fluorescence and spectral overlap of fluorochromes respectively. Cell viability and apoptosis were assessed by the 7AAD/AnnexinV/PI assay. In fact, cell viability was first assessed manually using the Trypan blue (Sigma-Aldrich) exclusion assay, and the results were then validated by the 7AAD method by flow cytometry. The telomerase activity [18 (link)] was assessed by real-time qPCR (LightCycler 2.0, Roche, Basel, Switzerland) using the Quantitative Telomerase Detection Kit (Cat#MT3012, Allied Biotech, Taipei, Taiwan) according to the manufacturer’s instructions.
Anderi R., Makdissy N., Azar A., Rizk F, & Hamade A. (2018). Cellular therapy with human autologous adipose-derived adult cells of stromal vascular fraction for alopecia areata. Stem Cell Research & Therapy, 9, 141.
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6

Flavivirus Infection Analysis in Cells

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The HC, trophoblasts, or U251 cells were infected as described above. Cells were fixed using CytoFix/Cytoperm and permeabilized according to the manufacturer's protocol (BD Biosciences, San Jose, CA), followed by staining with pan flavivirus 4G2 antibody (Millipore, Billerica, MA) and secondary staining with goat anti-mouse 488 (BD Biosciences). The HC were also stained with HLA-DR-VioBlue (Miltenyi Biotec) and DC-SIGN-APC (BD Biosciences), and 4G2+ cells, together with HLA-DR+ and DC-SIGN+ cells were quantified with flow cytometry (MacsQuant, Miltenyi Biotec). FACS plots were established based on forward and side scatter, doublet discrimination, and gates were set based on uninfected, stained control cell samples. Analysis was performed with MacsQuantify software (Miltenyi Biotec).
Gavegnano C., Bassit L.C., Cox B.D., Hsiao H.M., Johnson E.L., Suthar M., Chakraborty R, & Schinazi R.F. (2017). Jak Inhibitors Modulate Production of Replication-Competent Zika Virus in Human Hofbauer, Trophoblasts, and Neuroblastoma cells. Pathogens & Immunity, 2(2), 199-218.
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