Mouse t cell isolation kit

Manufactured by R&D Systems

The Mouse T cell isolation kit is a laboratory tool designed to separate T cells from mouse samples. It utilizes a negative selection method to enrich for T cells without directly labeling them.

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Lab products found in correlation

Anti mouse cd8 microbeads, by Miltenyi Biotec (1 mentions) 3h thymidine, by Hewlett-Packard (1 mentions) Scintillation counter, by Hewlett-Packard (1 mentions) 3h thymidine, by DuPont (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions) Cd3 cd28 dynabead, by Thermo Fisher Scientific (1 mentions)

4 protocols using mouse t cell isolation kit

T Cell Activation Assay with Dendritic Cells

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Untreated or XBP1/TRP2hsp70 or control DNA-transfected DC (6×10⁴) were co-cultured with syngeneic CD8⁺T cells (3×10⁵) isolated from splenocytes of naïve B6 mice using anti-mouse CD8 microbeads (Miltenyi Biotec) in 200 μl RPMI 1640 containing 10% (v/v) FBS for 3 days. ³H-thymidine (1 μCi/well; Du Pont/New England Nuclear, Boston, MA) was added during the last 16–18 hours of culture. ³H-thymidine incorporation was then measured using a scintillation counter (Packard, Meriden, CT) (23 (link)). In another set of experiments, untreated or XBP1/NeuEDhsp70 or control DNA-transfected DC (2x10⁴) were co-cultured with syngeneic naïve splenic T cells (1x10⁵) purified from naïve BALB/c mice using a mouse T cell isolation kit (R&D Systems) in 200 μl RPMI1640 media containing 10% (v/v) FBS for 5 days. IFN-γ in the culture supernatants were determined by ELISA.

Zhang Y., Chen G., Liu Z., Tian S., Zhang J., Carey C.D., Murphy K.M., Storkus W.J., Falo LD J.r, & You Z. (2015). Genetic Vaccines to Potentiate the Effective CD103+DC-mediated Cross-priming of Anti-tumor Immunity. Journal of immunology (Baltimore, Md. : 1950), 194(12), 5937-5947.

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Measuring T-cell Proliferation with MDSCs

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Isolation of CD4⁺CD3⁺ T cells was carried out with mouse T-cell isolation kit (R&D Systems) according to the manufacturer’s instruction. Isolated T cells were stained with 2 µM CSFE for 10 min and subsequently co-cultured with freshly isolated CD11b⁺Gr1⁺ MDSCs in different ratios (1:1; 2:1; 4:1, and 1:0) for 3 days. The fluorescence intensity of CFSE for each cell would halve after cell division. Number of cell division of each cell was analyzed by flow cytometry.

Chiu D.K., Tse A.P., Xu I.M., Di Cui J., Lai R.K., Li L.L., Koh H.Y., Tsang F.H., Wei L.L., Wong C.M., Ng I.O, & Wong C.C. (2017). Hypoxia inducible factor HIF-1 promotes myeloid-derived suppressor cells accumulation through ENTPD2/CD39L1 in hepatocellular carcinoma. Nature Communications, 8, 517.

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CFSE-based T cell division assay

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CD4⁺ CD3⁺ T cells were isolated using mouse T cell isolation kit (R&D system) according to the manufacturer’s instructions. Isolated T cells were stained with 2 μM 5,6-carboxyfluorescein diacetate succinimidyl ester (CFSE) for 10 min and mixed with freshly isolated CD11b⁺ Gr1⁺ MDSC at different ratios (1: 1.2, 1: 4, or 1: 10) and cultured for 3 days. The CFSE fluorescence intensity would reduce by 50% after the cells divided. Finally, the cell division was analyzed by flow cytometry.

Lei Y., Huang Y., Lin J., Sun S., Che K., Shen J., Liao J., Chen Y., Chen K., Lin Z, & Lin X. (2022). Mxi1 participates in the progression of lung cancer via the microRNA-300/KLF9/GADD34 Axis. Cell Death & Disease, 13(5), 425.

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Activated T Cell Proliferation Assay

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The isolation of T cells was carried out with a mouse T-cell isolation kit (R&D Systems, Minneapolis, MN) according to the manufacturer's instruction. T cells were stained with 2 mmol/L carboxyfluorescein succinimidyl ester (Thermo Fisher Scientific, Waltham, MA) for 10 minutes and in vitro activated with CD3/CD28 Dynabeads (Thermo Fisher Scientific) for 24 hours and subsequently cocultured with Hepa1-6 cells in 1:1 for 3 days. The number of cell divisions of each cell was analyzed by flow cytometry.

Chiu D.K., Yuen V.W., Cheu J.W., Wei L.L., Ting V., Fehlings M., Sumatoh H., Nardin A., Newell E.W., Ng I.O., Yau T.C., Wong C.M, & Wong C.C. (2020). Hepatocellular Carcinoma Cells Up-regulate PVRL1, Stabilizing PVR and Inhibiting the Cytotoxic T-Cell Response via TIGIT to Mediate Tumor Resistance to PD1 Inhibitors in Mice. Gastroenterology, 159(2).

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