T cell subsets of interest were flow-sorted from freshly isolated PBMCs at >98% purity using a modified FACSAria II (BD Biosciences). Cells were stained with the following reagents: (i) anti-CD3–APC-H7 (clone SK7), anti-CD14–V500 (clone M5E2), anti-CD19–V500 (clone HIB19), anti-CD28–APC (clone CD28.2), anti-CD45RA–PE (clone HI100), anti-CD57–FITC (clone NK-1), and anti-CCR7–PE-Cy7 (clone 3D12) from BD Biosciences; (ii) anti-CD4–PE-Cy5.5 (clone S3.5), anti-CD27–QD605 (clone CLB-27/1), and LIVE/DEAD Fixable Aqua from Thermo Fisher Scientific; (iii) anti-CD8–BV711 (clone RPA-T8), anti-CD95–PE-Cy5 (clone DX2), and anti-CD127–BV421 (clone A019D5) from BioLegend; and (iv) anti-CD45RO–ECD (clone UCHL1) from Beckman Coulter. Viable memory T cells were identified in the CD8+ lineage after gating out naive events on the basis of dual expression of CCR7 and CD45RA. Data were analyzed using FACSDiva software version 8.0 (BD Biosciences) and FlowJo software version 9.9.6 (FlowJo LLC).
Anti ccr7 pe cy7 clone 3d12
Manufactured by BD
The Anti-CCR7–PE-Cy7 (clone 3D12) is a fluorochrome-conjugated monoclonal antibody that can be used to detect the expression of the CCR7 receptor on cell surfaces. CCR7 is a chemokine receptor involved in lymphocyte trafficking and homing. This antibody can be used in flow cytometry applications to identify and characterize cell populations expressing CCR7.
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Lab products found in correlation
Live dead fixable aqua, by Thermo Fisher Scientific (2 mentions)
Anti cd45ro ecd clone uchl1, by Beckman Coulter (1 mentions)
Facsdiva software version 8, by BD (1 mentions)
Facsaria 2, by BD (1 mentions)
Anti cd8 apc cy7 clone sk1, by BD (1 mentions)
Anti cd45ra ecd clone 2h4ldh11ldb9, by Beckman Coulter (1 mentions)
Facsdiva software version 7, by BD (1 mentions)
Lsrfortessa, by BD (1 mentions)
2 protocols using anti ccr7 pe cy7 clone 3d12
1
Flow Cytometric Analysis of Memory T Cells
2
Multidimensional Phenotyping of PBMCs
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PBMCs were stained for surface markers using combinations of the following directly conjugated monoclonal antibodies: anti-CCR7–PE-Cy7 (clone 3D12; BD Biosciences), anti-CD3–BV605 (clone SK7; BD Biosciences), anti-CD8–APC-Cy7 (clone SK1; BD Biosciences), anti-CD4 BUV395 (clone SK3; BD Biosciences) anti-CD27–AF700 (clone O323; BioLegend), anti-CD45RA–ECD (clone 2H4LDH11LDB9; Beckman Coulter) and anti-CD95–FITC (clone DX2; BD Biosciences). Non-viable cells were eliminated from the analysis using LIVE/DEAD Fixable Aqua (Thermo Fisher Scientific). Samples were acquired using an LSR Fortessa (BD Biosciences). Data were analyzed using FACSDiva software version 7 (BD Biosciences) and/or FlowJo software version 10 (FlowJo LLC).
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