Ccr7 fitc g043h7

Manufactured by BioLegend

CCR7-FITC (G043H7) is a fluorescently-labeled antibody that recognizes the CCR7 receptor. CCR7 is a chemokine receptor that plays a role in lymphocyte trafficking and homing. The FITC (fluorescein isothiocyanate) fluorophore is conjugated to the antibody, allowing for detection and analysis of CCR7-expressing cells by flow cytometry.

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Lab products found in correlation

Cd161 pe and cd161 apc, by Miltenyi Biotec (2 mentions) Vδ2 percp cy5.5 b6, by BioLegend (2 mentions) Cd62l pe dreg 56, by BioLegend (2 mentions) Cd3 efluor450 okt3, by Thermo Fisher Scientific (2 mentions) Cd56 pe cy7 hcd56, by BioLegend (2 mentions) Live dead fixable near ir dead cell stain kit, by Thermo Fisher Scientific (2 mentions) Macsquant analyzer 10, by Miltenyi Biotec (2 mentions) Cd45 percp 2d1, by BioLegend (1 mentions) Facsaria 2 cell sorter, by BD (1 mentions) Cd8 viogreen, by Miltenyi Biotec (1 mentions) Benzonase endonuclease, by Merck Group (1 mentions) Live dead aqua dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Cd27 bv711, by BioLegend (1 mentions) Cd28 bv421 cd28.2, by BioLegend (1 mentions)

4 protocols using ccr7 fitc g043h7

Quantifying Immune Cell Phenotypes

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The percentages of GFP⁺ HeLa cells, 293T cells, U937 cells, and monocytes were evaluated by suspending the cells in PBS and fixing them with 2% paraformaldehyde for 10 min prior to flow cytometry analysis using a FACSAria II cell sorter (BD Biosciences). Intact cells were gated using forward scatterplots versus side scatterplots, and further gating for KI cells was performed based on the presence of control samples expressing GFP using fluorescein isothiocyanate (FITC) versus side scatterplots. The number of GFP⁺ cells in control samples ranged from 0% to 1%.
At the time of harvest, PBMCs were stained with antibodies against the following proteins: CD45-PerCp (2D1), CD4-phycoerythrin (PE)/Cy7 (OKT4), CD8-allophycocyanin (APC)/Cy7 (SK1), CD45RA-APC (HI100), CCR7-FITC (G043H7), CD38-APC (HIT2), HLA-DR-PE (L243), PD-1-PE (EH12.2H7), PD-L1-APC (29E.2A3), and CTLA-4-FITC (A3.4H3.H12) (all from BioLegend). The lymphocyte gate was based on CD45. Among lymphocytes, T cells were sub-gated based on their expression of CD4 or CD8. Memory subsets of CD4⁺ and CD8⁺ T cells were defined by the expression of CCR7 and CD45RA, respectively. The activation status was determined based on CD38 and HLA-DR expression, whereas the inhibitory status was determined based on PD-1, PD-L1, and CTLA-4. The gates for activation markers and inhibitory markers were determined using isotype control antibodies.

Wang B., Zuo J., Kang W., Wei Q., Li J., Wang C., Liu Z., Lu Y., Zhuang Y., Dang B., Liu Q., Kang W, & Sun Y. (2018). Generation of Hutat2:Fc Knockin Primary Human Monocytes Using CRISPR/Cas9. Molecular Therapy. Nucleic Acids, 11, 130-141.

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Multiparameter Flow Cytometry Staining

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Cells were stained with a viability dye (Live/Dead fixable near-IR dead cell stain kit, Invitrogen) and fluorochrome-labelled antibodies for 30 minutes at 4°C. The following antibodies were used: CD3-eFluor450 (OKT3, eBioscience), CD8-VioGreen (BW135/80, Miltenyi), γδTCR-FITC (5A6.E9, Invitrogen), CD161-PE and CD161-APC (191B8, Miltenyi), Vδ2-PerCP-Cy5.5 (B6, BioLegend), CD56-PE-Cy7 (HCD56, BioLegend), Vα7.2-APC and Vα7.2-PerCP-Cy5.5 (3C10, BioLegend), CCR7-FITC (G043H7), CD62L-PE (DREG-56, BioLegend). Data was acquired by a MACSQuant Analyzer 10 (Miltenyi).

Hagel J.P., Bennett K., Buffa F., Klenerman P., Willberg C.B, & Powell K. (2021). Defining T Cell Subsets in Human Tonsils Using ChipCytometry. The Journal of Immunology Author Choice, 206(12), 3073-3082.

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Comprehensive T Cell Phenotyping Protocol

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T cell phenotyping was conducted in Oxford for all 16 UK volunteers and 27 Senegalese volunteers for whom there were cryopreserved cells available. PBMCs were thawed and rested for 2 h at 37°C in 2.5 µl/ml Benzonase Endonuclease (E1014-25KU; Sigma-Aldrich). One to two million PBMCs for each individual were stained in 50 µl in 96-well plates. Cells were incubated for 20 min at room temperature (RT) with the Live/Dead Aqua Dead Cell Stain Kit (Invitrogen) then surface stained at RT with CD14-eF506 (61D3, 1/50; eBioscience), CD19-eF506 (HIB19, 1/50; eBioscience), CD45RA-eV605 (HI100, 1/50; eBioscience), CD27-BV711 (O323, 3/50; BioLegend), CD28-BV421 (CD28.2, 1/50; BioLegend), CD4-APC (RPA-T4, 1/50; eBioscience), CD3-AF700 (UCHT1, 1/50; eBioscience), CD8-APC-AF780 (RPA-T8, 3/50; eBioscience), CCR7-FITC (G043H7, 1/50; BioLegend), CD57-PerCP-Cy5.5 (HNK-1, 1/50; BioLegend), and KLRG1-PE (14C2A07, 3/50; BioLegend). Cells were acquired immediately using a BD LSRII. Data analysis was conducted in FlowJo version 10.1 (Treestar Inc). Gating strategy is shown in Fig. S1.

Bowyer G., Sharpe H., Venkatraman N., Ndiaye P.B., Wade D., Brenner N., Mentzer A., Mair C., Waterboer T., Lambe T., Dieye T., Mboup S., Hill A.V, & Ewer K.J. (2020). Reduced Ebola vaccine responses in CMV+ young adults is associated with expansion of CD57+KLRG1+ T cells. The Journal of Experimental Medicine, 217(7), e20200004.

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Immunophenotyping of T Cell Subsets

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Cells were stained with a viability dye (LIVE/DEAD Fixable Near-IR Dead Cell Stain Kit; Invitrogen) and fluorochrome-labeled Abs for 30 min at 4°C. The following Abs were used: CD3–eFluor 450 (OKT3; eBioscience), CD8-VioGreen (BW135/80; Miltenyi Biotec), γδTCR-FITC (5A6.E9; Invitrogen), CD161-PE and CD161-APC (191B8; Miltenyi Biotec), Vδ2-PerCP-Cy5.5 (B6; BioLegend), CD56–PE-Cy7 (HCD56; BioLegend), Vα7.2-APC and Vα7.2-PerCP-Cy5.5 (3C10; BioLegend), CCR7-FITC (G043H7), and CD62L-PE (DREG-56; BioLegend). Data were acquired by a MACSQuant Analyzer 10 (Miltenyi Biotec).

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