WT BV173 cells were co-cultured with either sTCR+ or sTCR+CoCAR3+ T cells at an E:T ratio of 1:5 for two rounds of serial co-culture as described above. T cells were collected, mixed with BV173 cells at a ratio of 1:5 for 24 hours, and then separated using CD4 and CD8 MACS beads (Miltenyi Biotech, Germany. #130–045-101 and 130–045-201). Mitochondrial function was determined using Agilent Seahorse XFe96 Extracellular Flux Assay Kits (Agilent, Santa Clara, CA, Cat # 102416-), and analysis was conducted by the Baylor College of Medicine Mouse Metabolic and Phenotyping Core. Data were analyzed using the Seahorse Wave software (ECAR and OCR / Agilent).
Cd4 and cd8 macs beads
Manufactured by Miltenyi Biotec
Sourced in Germany
CD4 and CD8 MACS beads are magnetic particles used for the separation and isolation of CD4-positive and CD8-positive cells from complex biological samples, such as peripheral blood mononuclear cells (PBMCs) or single-cell suspensions. The beads are coated with antibodies specific to the CD4 or CD8 surface markers, allowing for the efficient and gentle isolation of the target cell populations.
Automatically generated - may contain errors
2 protocols using cd4 and cd8 macs beads
1
T Cell Metabolic Profiling in Co-Culture
2
Mitochondrial Function Assay for sTCR+ and sTCR+CoCAR3+ T Cells
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WT BV173 cells were cocultured with either sTCR+ or sTCR+CoCAR3+ T cells at an E:T ratio of 1:5 for two rounds of serial coculture as described above. T cells were collected, mixed with BV173 cells at a ratio of 1:5 for 24 hours, and then separated using CD4 and CD8 MACS beads (Miltenyi Biotec, Germany, #130-045-101 and 130-045-201). Mitochondrial function was determined using Agilent Seahorse XFe96 Extracellular Flux Assay Kits (Agilent, catalog no. 102416), and analysis was conducted by the Baylor College of Medicine Mouse Metabolic and Phenotyping Core. Data were analyzed using the Seahorse Wave software (ECAR and OCR/Agilent).
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