Blood naïve T cells were purified after ficoll gradient using the Naïve Pan T Cell Isolation Kit (Miltenyi Biotec) and stained with Cell Proliferation Dye eFluor-670 (eBioscience) as recommended by the manufacturer. Purified mucosal CD1a+ or CD14+ were plated with naïve T cells (1:15) in round-bottom 96-well plates, in Xvivo 15 media (Invitrogen) supplemented with 10% human AB serum (Valley Biomedical). After 6 days in culture, proliferation of T cells was assessed by flow cytometry after staining with zombie yellow dye (Biolegend) and CD3-APC-Cy7 (Tonbo). As a control, blood monocyte-derived DC were generated in vitro with IL-4 and GM-CSF66 (link) after CD14+ selection as previously described16 (link).
Cd3 apc cy7
Manufactured by Cytek Biosciences
Sourced in United States
CD3-APC-Cy7 is a fluorochrome-conjugated monoclonal antibody that binds to the CD3 antigen, a component of the T cell receptor complex. It is used for the identification and enumeration of T cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Zombie yellow dye, by BioLegend (2 mentions)
Cell proliferation dye efluor 670, by Thermo Fisher Scientific (2 mentions)
Na ve pan t cell isolation kit, by Miltenyi Biotec (2 mentions)
Xvivo 15 media, by Thermo Fisher Scientific (2 mentions)
Human ab serum, by Valley Biomedical (2 mentions)
Cd8 fitc, by Cytek Biosciences (2 mentions)
Apc conjugated streptavidin, by BioLegend (1 mentions)
Pe conjugated streptavidin, by BioLegend (1 mentions)
Penicillin streptomycin, by Welgene (1 mentions)
Cd3 apc, by BioLegend (1 mentions)
Cd11c percp cy5, by BioLegend (1 mentions)
Cd4 pe, by Thermo Fisher Scientific (1 mentions)
Pd l1 pe cy7, by BD (1 mentions)
Cd163 apc, by BioLegend (1 mentions)
Cd45 violetfluor 450, by Cytek Biosciences (1 mentions)
Pd 1 percp cy5, by BD (1 mentions)
Cd11b pe, by Cytek Biosciences (1 mentions)
Macsquant flow cytometer, by Miltenyi Biotec (1 mentions)
4 protocols using cd3 apc cy7
1
Naïve T Cell Proliferation Assay with Mucosal APCs
2
Murine Splenocyte Immune Profiling
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Mouse splenocytes were cultured in a complete RPMI-1640 medium (GenDEPOT, Katy, TX, USA) supplemented with fetal bovine serum (10%, Hyclone, South Logan, UT, USA) and 1% penicillin/streptomycin (Welgene, Gyeongsan, Korea). Anti-mouse TCR-β-PE, CD45.1-Percp Cy5.5, CD3-APC Cy7, CD8-FITC, CD44-PE, and IFN-γ-FITC antibodies as well as CFSE cell proliferation tracing dye were purchased from Tonbo Bioscience (San Diego, CA, USA). Anti-mouse TNF-α-PE Cy7, APC-conjugated streptavidin, and PE-conjugated streptavidin were purchased from Biolegend (San Diego, CA, USA). Gp33-41 class I pMHC tetramer was provided by the NIH Tetramer Core Facility (Atlanta, GA, USA).
3
Naïve T Cell Proliferation Assay with Mucosal APCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Blood naïve T cells were purified after ficoll gradient using the Naïve Pan T Cell Isolation Kit (Miltenyi Biotec) and stained with Cell Proliferation Dye eFluor-670 (eBioscience) as recommended by the manufacturer. Purified mucosal CD1a+ or CD14+ were plated with naïve T cells (1:15) in round-bottom 96-well plates, in Xvivo 15 media (Invitrogen) supplemented with 10% human AB serum (Valley Biomedical). After 6 days in culture, proliferation of T cells was assessed by flow cytometry after staining with zombie yellow dye (Biolegend) and CD3-APC-Cy7 (Tonbo). As a control, blood monocyte-derived DC were generated in vitro with IL-4 and GM-CSF66 (link) after CD14+ selection as previously described16 (link).
4
Multicolor Flow Cytometry Analysis
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