Abs against Thy1.2 (53–2.1), CD24 (M1/69), CD25 (PC61), CD44 (IM7), TCR-δ (GL3), TCR-β (H57-597), CD4 (GK1.5), CD8α (53–6.7), CD27 (LG.7F9), CD45RB (C363-16A), CD73 (Ty/11.8), Vγ2 (UC3-10A6), Vγ3 (536), and NK1.1 (PK136) were purchased from eBioscience, BD, or BioLegend. Anti-Vγ1.1 antibody was provided by R. O’Brien (National Jewish Medical Center, Denver, CO). Propidium iodide gating was included for dead cell exclusion. Data were collected on either a LSRII or FACSVantage SE (BD) and analyzed using FlowJo Software (Tree Star). For isolation of thymic γδ T cell populations, thymocytes were harvested from 6–7-wk-old C57BL/6 mice, depleted with anti-CD4 and anti-CD8 magnetic beads (Miltenyi Biotec), and isolated by cell sorting. A dump gate (NK1.1, B220, Ter11, Gr-1, and CD11b) was included. Intracellular TCR chains were detected by blocking surface TCR using unlabeled Ab, fixing for 10 min with 1% paraformaldehyde at room temperature, permeabilization with saponin and NP-40 detergents, and then staining with fluorochrome-conjugated anti–TCR-β and TCR-δ Ab.
Anti cd4 and anti cd8 magnetic beads
Manufactured by Miltenyi Biotec
Anti-CD4 and anti-CD8 magnetic beads are laboratory reagents used for the isolation and enrichment of CD4+ and CD8+ T cells from biological samples. These beads are coated with antibodies specific to the CD4 and CD8 surface proteins, allowing for the targeted separation and purification of the respective T cell subsets.
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Lab products found in correlation
4 protocols using anti cd4 and anti cd8 magnetic beads
1
Comprehensive Thymic γδ T Cell Phenotyping
2
Single-cell RNA-seq of Murine Hepatic and Thymic γδ T Cells
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Hepatic and thymic MNCs were harvested from B6 mice (8 weeks old). Thymic cell suspensions were enriched for γδ T cells by magnetic bead negative selection using anti-CD4 and anti-CD8 magnetic beads (Miltenyi Biotec). Cells were stained with anti-CD3e and anti-TCRγ/δ antibodies, as described above. After surface staining, hepatic or thymic γδ T cells were sorted using a MoFlo Astrios EQ. After sorting, the cells were washed and maintained on ice. ScRNA-Seq libraries were prepared using the 10× Genomics Chromium single-cell 3’ v2 kit, according to standard protocol provided by CapitalBio Technology Co., Ltd. (Beijing, China). RNA-seq was performed on an Illumina HiSeq X Ten system using 150 bp paired-end sequencing (PE150). All sequencing data have been submitted to the National Center for Biotechnology Information Gene Expression Omnibus (NCBI GEO) depository under the accession number GEO: GSE164106.
3
CD4+ Treg-Mediated Suppression of Naive T Cells
4
CD4+ Treg-Mediated Suppression of Naive T Cells
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Sorted CD4+Foxp3-GFP+ Treg cells were co-cultured with sorted CD4+CD62L+Foxp3-GFP− naïve T cells that were stimulated with anti-CD3ε ε (1 μg/ml) in the presence of irradiated antigen presenting cells (APCs) for 3 days at 37 °C, 5% CO2. APCs, isolated from wild-type spleens after T cell depletion using anti-CD4 and anti-CD8 magnetic beads (Miltenyi Biotec), or Rag1−/− spleens were irradiated prior to use.
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