Pan t cells isolation kit

Manufactured by Miltenyi Biotec

Sourced in United States

The Pan T-cell Isolation Kit is a laboratory product designed to isolate and enrich T-cells from various sample types. It utilizes a magnetic bead-based separation method to selectively deplete non-T-cells, resulting in a highly purified T-cell population.

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Lab products found in correlation

Cd14 immunomagnetic beads, by Miltenyi Biotec (1 mentions) Appropriate isolation kits, by Miltenyi Biotec (1 mentions) Cd8 t cell isolation kit, by Miltenyi Biotec (1 mentions) Zombie aqua fixable viability kit, by BioLegend (1 mentions)

Close spelling variants of this product

Pan T Cell Isolation Kit (429 citations) T cell isolation kit (44 citations) Pan-T isolation kit (16 citations) Isolation kits (10 citations) Cell isolation kits (5 citations) Pan-T cells (4 citations) Pan T cells isolation kit II (4 citations) Human Pan T cells isolation kit (3 citations)

6 protocols using pan t cells isolation kit

Leukocyte Cytokine-Induced MMP-9 Secretion

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Peripheral Blood Mononuclear Cells (PBMCs) and PMN from healthy donors were obtained by density-gradient centrifugation from EDTA-treated whole blood using a density gradient medium (Granulosep, Eurobio-Abcys). Monocytes were purified from PBMCs by positive selection using CD14 immunomagnetic beads (MACS; Miltenyi Biotech) according to manufacturer instructions. Flow-through was then used to isolate T-lymphocyte by negative selection using the Pan T-cells isolation kit from Miltenyi (MACS).
Isolated leukocytes from healthy donors were then cultured in FBS-free medium and stimulated overnight (lymphocytes and monocytes) or for 1 hour (PMN) by IL-17 (200 pg/mL), IL-23 (100 pg/mL), or both cytokines. Culture media were harvested and analyzed for MMP-9 secretion by gel zymography.

Plée J., Le Jan S., Giustiniani J., Barbe C., Joly P., Bedane C., Vabres P., Truchetet F., Aubin F., Antonicelli F, & Bernard P. (2015). Integrating longitudinal serum IL-17 and IL-23 follow-up, along with autoantibodies variation, contributes to predict bullous pemphigoid outcome. Scientific Reports, 5, 18001.

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Allogeneic Bone Marrow Transplant in Irradiated Mice

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Recipient B6D2F1 female mice received a 10-Gy irradiation followed by retro-orbital infusion of 5 × 10⁶ bone marrow cells (C57BL/6J) + 1 × 10⁶ CD3⁺ T cells from WT littermate or Suv39h1-KO male mice + 2 × 10⁴ mastocytoma cells P815-GFP derived from DBA/2 (H-2^d) mice (gift from Dr. B. Salomon). Total splenocytes and lymph nodes suspension cells were prepared for CD3⁺ T cells enrichment using pan T cells isolation kit (Miltenyi). Bone marrow suspension were prepared using leg bones. Control groups were constituted of irradiated mice receiving only bone marrow or bone marrow and P815-GFP cells. GVHD and Leukemia was evaluated two times per week. GVHD was evaluated as previously described^{48 (link),49 (link)}, and leukemia by flow cytometry (GFP⁺ H-2K^{d +}cells) and tumor bearing eyes.

Niborski L.L., Gueguen P., Ye M., Thiolat A., Ramos R.N., Caudana P., Denizeau J., Colombeau L., Rodriguez R., Goudot C., Luccarini J.M., Soudé A., Bournique B., Broqua P., Pace L., Baulande S., Sedlik C., Quivy J.P., Almouzni G., Cohen J.L., Zueva E., Waterfall J.J., Amigorena S, & Piaggio E. (2022). CD8+T cell responsiveness to anti-PD-1 is epigenetically regulated by Suv39h1 in melanomas. Nature Communications, 13, 3739.

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Activation of Lymph Node T Cells

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T cells were purified from LN with Pan T cells Isolation Kit (Miltenyi Biotec). LN T cells were stimulated with or without plate-bound anti-TCRβ (1 μg/ml) and anti-CD28 (1 μg/ml) antibodies for 24 hours at 37 °C. Cultured cells were stained and analyzed by flow cytometry for CD40L and CD69 expression.

Shinzawa M., Moseman E.A., Gossa S., Mano Y., Bhattacharya A., Guinter T., Alag A., Chen X., Cam M., McGavern D.B., Erman B, & Singer A. (2022). Reversal of the T cell immune system reveals the molecular basis for T cell lineage fate determination in the thymus. Nature Immunology, 23(5), 731-742.

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Isolation of T cells from Mice

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For T cells isolation, mice were sacrificed at the designated days of experiments to remove spleen and lymph nodes. The splenocytes were obtained by macerating spleens with a plunger and passing the tissue homogenates through a 70μm cell strainer. Cells were washed 2-3 times by centrifugation at 500g at 4°C for 5 min with incomplete RPMI-1640 to obtain a single-cell suspension. Pan-T cells were isolated using Pan-T cells isolation kit (Miltenyi Biotec, USA) using magnetic columns as per the manufacturer’s instructions. As and when required, the CD4⁺ and CD8⁺ T cells were isolated by positive selection using appropriate isolation kits (Miltenyi Biotec, USA). For isolation of lymph nodes (LN) lymphocytes, the brachial, axillary, and inguinal nodes were harvested and pooled together and the cells were isolated by mechanical maceration as described above.

Singh R., Anand A., Rawat A.K., Saini S., Mahapatra B., Singh N.K., Mishra A.K., Singh S., Singh N., Kishore D., Kumar V., Das P, & Singh R.K. (2022). CD300a Receptor Blocking Enhances Early Clearance of Leishmania donovani From Its Mammalian Host Through Modulation of Effector Functions of Phagocytic and Antigen Experienced T Cells. Frontiers in Immunology, 12, 793611.

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Isolation and Activation of Tumor-Reactive T Cells

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A Pan-T cells Isolation kit (Miltenyi Biotec) and a CD8 T cell Isolation kit (Miltenyi Biotec) were used to isolate T cells from mouse spleen. T cells were primed by different groups of p13nsEV treatment for 5 days. After priming, the p13nsEV was washed away, and T cells were cocultured with tumor cells at 10:1 (tumor cell:T cell) for 2 days before being tested for interferon-γ (IFN-γ) expression by FC. For the T cell cytotoxicity assay, the p13nsEV-treated T cells were then cocultured with GFP-labeled cancer cells, which is the same source of tumor lysate to generate the p13nsEV. After 5 days of coculture, the death of cancer cells was examined by staining cells using the Zombie Aqua Fixable Viability Kit (BioLegend) followed by FC.

Wu K., Lyu F., Wu S.Y., Sharma S., Deshpande R.P., Tyagi A., Zhao D., Xing F., Singh R, & Watabe K. (2023). Engineering an active immunotherapy for personalized cancer treatment and prevention of recurrence. Science advances, 9(17).

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Transfer of Splenic T Cells in Mice

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Splenic T cells from C57BL/6 (CD45.1), WT (CD45.2) and Ahrr^−/− (CD45.2) mice were isolated using Pan T Cells Isolation Kit (Miltenyi Biotech) according to manufacturer’s instruction. Both CD45.1 and CD45.2 cells were mixed together in 1:1 ratio. Rag1^−/− mice were injected with 10⁶ cells intravenously. Frequencies of CD45.1 vs CD45.2 in different subsets of IEL were analyzed after 8 weeks in the small intestine of recipient mice.

Panda S.K., Peng V., Sudan R., Antonova A.U., Di Luccia B., Ohara T.E., Fachi J.L., Grajales-Reyes G., Jaeger N., Trsan T., Gilfillan S., Cella M, & Colonna M. (2023). Repression of the Aryl Hydrocarbon Receptor Prevents Oxidative Stress and Ferroptosis of Intestinal Intraepithelial Lymphocytes. Immunity, 56(4), 797-812.e4.

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