To assess the cytotoxicity of (1 ), (2 ), and (5 ) in human tonsillar tissues after 12 days of culture, cells isolated from untreated tissue blocks and from those treated with compounds were stained with combinations of the following fluorescence- labeled monoclonal antibodies: anti– CD3-QD605, anti–CD4-QD655, anti–CD8-QD705, anti–CD25-APC, anti–CD38-PE, anti–HLA-DR-APC-Cy7, anti–CXCR4-Brilliant violet 421, anti–CCR5-PR-Cy5 anti–CD45RA-FITC, and anti–CCR7-PE-Cy7 (Caltag Laboratories; Biolegend). Detection and enumeration of HIV-1– infected cells were performed with intracellular staining by means of anti–p24-PE (Beckman Coulter). Data were acquired and analyzed as described elsewhere (Grivel and Margolis, 2009 ). We quantified cell depletion using Trucount beads (Becton Dickinson) for volumetric control and normalized cell numbers by tissue-block weights.
Anti hla dr apc cy7
Manufactured by BioLegend
Anti–HLA-DR-APC-Cy7 is a fluorescently-labeled antibody that binds to the HLA-DR antigen, which is expressed on the surface of antigen-presenting cells. This product can be used for the detection and analysis of HLA-DR-positive cells in flow cytometry applications.
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Lab products found in correlation
Trucount beads, by BD (1 mentions)
Anti cd25 apc, by BioLegend (1 mentions)
Anti ccr7 pe cy7, by BioLegend (1 mentions)
Anti p24 pe, by Beckman Coulter (1 mentions)
Anti cd38 pe, by BioLegend (1 mentions)
Positive and negative compensation beads, by BD (1 mentions)
Pe cy7 anti cd83, by BioLegend (1 mentions)
Anti pd l1 brilliant violet 421, by BioLegend (1 mentions)
Fitc anti cd80, by BioLegend (1 mentions)
Human trustain fcx, by BioLegend (1 mentions)
Anti cd163 pe, by BioLegend (1 mentions)
Zombie violet, by BioLegend (1 mentions)
Lyric flow cytometer, by BD (1 mentions)
Anti pd l1 apc, by BioLegend (1 mentions)
Ifn γ, by R&D Systems (1 mentions)
Accutase, by Merck Group (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Interleukin 4 (il 4), by R&D Systems (1 mentions)
3 protocols using anti hla dr apc cy7
1
Cytotoxicity Assessment of Compounds in Tonsillar Tissues
2
Comprehensive Dendritic Cell Immunophenotyping
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Dendritic cells were stained with the following antibodies: anti-CD83 Pe-Cy7, anti-PD-L1 Brilliant Violet 421, anti-CD11c Brilliant Violet 510, anti-CD54 allophycocyanin (APC), and anti-HLA-DR APC-Cy7 (Biolegend) for 15 min at room temperature in the dark. Samples were washed in PBA buffer (PBS containing 2% FCS and 0.01% v/v sodium azide) and acquired on DAKO CyAn flow cytometer (Beckman Coulter).Compensation was performed using positive and negative compensation beads (BD Biosciences) and data were analysed with FlowJo software (Tree Star Inc.) version 543 (link),48 (link).
3
Macrophage Polarization by Cytokines and RABV
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Mature macrophages (n = 6 donors) were stimulated for 48 hours with complete medium containing IFN-g (20 ng/mL, R&D Systems) and LPS (100 ng/mL, Sigma Aldrich), with IL-4 (20 ng/mL, R&D Systems), or with IL-1 (20 ng/mL R&D Systems), to induce the M1, M2a or M2c phenotype, respectively. To investigate the effect of rabies virus on macrophage polarization, macrophages were stimulated with complete medium containing RABV (MOI of 10). Expression of macrophage phenotypical markers was investigated by flow cytometry. Non-polarized macrophages, cultured for 48 hours with complete medium without additional cytokines, were taken along as controls for each donor.
Macrophages were dissociated from the wells using Accutase (Merck Millipore) and were washed twice with PBS before staining for 30 minutes with the fixable viability dye ZombieViolet (Biolegend). Cells were fixed with 4 % PFA for 15 minutes, and after Fc receptor blocking with Human TruStain FcX (Biolegend), cells were stained with the following antibodies in FACS buffer (PBS with 2 % fetal calf serum, 0.2 mM EDTA, 0.01% sodium azide): anti-CD80-FITC, anti-HLA-DR-APC-Cy7, anti-PD-L1-APC, anti-CD163-PE, anti-CD200R-PE-Cy7 and anti-CD206 (BV786) (All Biolegend). Mean fluorescent intensities were quantified by flow cytometry using a BD Lyric flow cytometer (BD Biosciences) and the data were analyzed using FlowJo V10.6.2.
Macrophages were dissociated from the wells using Accutase (Merck Millipore) and were washed twice with PBS before staining for 30 minutes with the fixable viability dye ZombieViolet (Biolegend). Cells were fixed with 4 % PFA for 15 minutes, and after Fc receptor blocking with Human TruStain FcX (Biolegend), cells were stained with the following antibodies in FACS buffer (PBS with 2 % fetal calf serum, 0.2 mM EDTA, 0.01% sodium azide): anti-CD80-FITC, anti-HLA-DR-APC-Cy7, anti-PD-L1-APC, anti-CD163-PE, anti-CD200R-PE-Cy7 and anti-CD206 (BV786) (All Biolegend). Mean fluorescent intensities were quantified by flow cytometry using a BD Lyric flow cytometer (BD Biosciences) and the data were analyzed using FlowJo V10.6.2.
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