Cells were surface stained, fixed, and refrigerated at 4°C overnight, followed by flow cytometry using the BD LSRII instrument according to manufacturer’s protocols (BD Biosciences, Carlsbad, CA, USA). An 11-color flow panel of cross-reactive antibodies was used: CD3 Alexa700, CD4 BD-V500, CD8 BD-V450, CD20 FITC, CCR5 PE, CXCR4 PE-CF594, CCR7 PE-Cy7, CD45RA PE-Cy5, CD14 APC, CD69 APC-H7, and HLA-DR BV605 (BD Biosciences). Samples were analyzed using FlowJo (TreeStar Inc, Ashland, OR, USA), and results graphed using GraphPad Prism 5 (Graphpad Software Inc, La Jolla, CA, USA).
Cd69 apc h7
Manufactured by BD
CD69 APC-H7 is a fluorochrome-conjugated antibody that binds to the CD69 protein, which is a cell surface marker expressed on activated T cells, B cells, and natural killer cells. It can be used in flow cytometry analysis to identify and quantify activated immune cells.
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Lab products found in correlation
Ccr7 pe cy7, by BD (1 mentions)
Cd3 alexa 700, by BD (1 mentions)
Lsrii instrument, by BD (1 mentions)
Cd45ra pe cy5, by BD (1 mentions)
Ccr5 pe, by BD (1 mentions)
Cd20 fitc, by BD (1 mentions)
Hla dr bv605, by BD (1 mentions)
Cd14 apc, by BD (1 mentions)
Prism 5, by GraphPad (1 mentions)
Anti pd 1 pe, by BD (1 mentions)
2 protocols using cd69 apc h7
1
Multiparametric Flow Cytometry Analysis
2
Allogeneic T-cell Proliferation Assay
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The functional activity of DCs was first evaluated through their ability to stimulate the proliferation of normal allogeneic T lymphocytes. DCs from six different donors were cocultured with allogeneic T lymphocytes (previously marked with carboxyfluorescein succinyl ester (CFSE)) in flat-bottomed 96-well plates in a 1:10 (104:105) DCs: lymphocyte ratio. Cells were harvested five days later, and the lymphocyte proliferation was analyzed by flow cytometry based on the dilution of CFSE in the replicant cells. We also analyzed the expression of PD-1 and CD69 on CD3+ cells using anti-PD-1-PE and CD69-APC-H7 (BD Pharmingen).
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