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Anti cd8 macs beads

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Anti-CD8 MACS beads are magnetic microbeads coated with antibodies that specifically bind to the CD8 molecule expressed on the surface of certain immune cells, such as cytotoxic T cells. These beads can be used to isolate and enrich CD8-positive cells from complex cell samples.

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Lab products found in correlation

Interleukin 2 (il 2), by Thermo Fisher Scientific (2 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions) Facsaria, by BD (1 mentions) Ls column, by Miltenyi Biotec (1 mentions) Anti cd4, by Miltenyi Biotec (1 mentions) Heat inactivated, by Merck Group (1 mentions) Interleukin 7 (il 7), by Thermo Fisher Scientific (1 mentions) Il 15, by R&D Systems (1 mentions)

Close spelling variants of this product

CD8 MACS beads Anti-CD8 beads MACS beads Anti-CD8

6 protocols using anti cd8 macs beads

1

T Cell Transfer for Tumor Immunotherapy

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For T cell transfer, T circulating CD8+ T cells from the spleen, or T resident memory cells from the skin inguinal adipose tissue of survivor mice were isolated using anti-CD8 MACS beads (Miltenyi Biotec). Enriched cells were then sorted as live CD45+CD3+CD8+ cells using FACS ARIA. Purity post-sorting was > 97%. Sorted T cells were then stimulated for 24 h in 24-well plates (~2 × 106 cells/well) coated with anti-CD3 (4 μg/ml) and anti-CD28 (4 μg/ml) in the presence of IL-2 (Peprotech, 50ng/ml). 5.×105 of activated splenic or skin inguinal adipose CD8+T cells were transferred by i.v. or intra dermal (i.d.) injection respectively on the right flank into naïve recipient mice 7 days prior tumor challenge. Some mice received both T cell subsets 7 days prior tumor challenge.
Zhivaki D., Borriello F., Chow O.A., Doran B., Fleming I., Theisen D.J., Pallis P., Shalek A.K., Sokol C.L., Zanoni I, & Kagan J.C. (2020). Inflammasomes within Hyperactive Murine Dendritic Cells Stimulate Long-Lived T Cell-Mediated Anti-tumor Immunity. Cell reports, 33(7), 108381.
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2

Isolation and Characterization of Mouse CD8+ T Cells

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CD8+ T cells from influenza-infected C57BL/6 mice or OT-I mice were obtained by depletion of CD11c+ cells with anti-CD11c MACS beads followed by positive selection with anti-CD8 MACS beads (Miltenyi Biotec). All T cell preparations were more than 95% pure. In some experiments, CD8+ T cells were labeled for 10 min at 37 °C with 5 µM CFSE (Molecular Probes).
CD8+CD44hi memory T cells were sorted from spleens of C57BL/6 or Cd40−/− mice using a FACSAria (BD Biosciences) after positive selection with anti-CD8 MACS beads. Cell numbers were normalized to the concentration of antigen-specific T cells and 4 × 104 CD8+CD44hi DbNP+ or CD8+CD44hi DbPA+ T cells were transferred intravenously into naïve C57BL/6, Cd40−/− or CD45.1 recipient mice. DCs were enriched from pooled mLNs of C57BL/6, Cd40−/− or Cd154−/− with anti-CD11c MACS beads. In some experiments, DC subsets were sorted with a FACSAria. All sorted DC subsets were more than 95% pure.
Ballesteros-Tato A., León B., Lee B.O., Lund F.E, & Randall T.D. (2014). Epitope-specific regulation of memory programming by differential duration of antigen presentation to influenza-specific CD8+ T cells. Immunity, 41(1), 127-140.
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3

Adoptive Transfer of CFSE-Labeled OTI T Cells

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CD8+ T cells from spleens and LNs of OTI × B6.SJL-Ptprca mice were positively selected using anti-CD8+ MACS Beads (Miltenyi Biotec) and labeled with CFSE as described (32 (link)). Purity was >87% as assessed by positive staining with anti-CD8α, -Vα2, and -Vβ5 antibodies. About 2 × 106 cells were injected i.v. into C57BL/6J or CD11c-DTR BM chimeric hosts bearing established B16.OVA tumors. OTI proliferation was assessed in tumor-draining LNs 6 days later.
Kuhn S., Yang J, & Ronchese F. (2015). Monocyte-Derived Dendritic Cells Are Essential for CD8+ T Cell Activation and Antitumor Responses After Local Immunotherapy. Frontiers in Immunology, 6, 584.
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4

Adoptive Transfer of OVA-Specific T Cells

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CD8+ T and CD4+ T cells from the spleens of naïve B6.OTI or B6.CD45.1+ OTII mice were isolated using anti-CD8 MACS beads and LS columns (Miltenyi Biotec) according to the manufacturer´s instructions for CD8+ T cells, or using negative selection MagCellect kits (R&D systems) for CD4+ T cells. For CD4+ OTII T cell transfer experiments, cells were stimulated with plate-bound anti-CD3 (clone 145-2C11, 5µg/mL) and anti-CD28 (clone 37.51, 10µg/mL) and polarized in type 1 conditions (anti-IL4 (clone 11B11), 10µg/mL) and IL-12 (R&D catalog number 419ML-010, 5ng/mL) in the absence (Ctrl) or presence of DON for three days. In the indicated experiments (25 × 103) of OTI cells or (25 × 104) of OTII cells were intravenously injected into the indicated recipient mice. Recipient mice of OTI cells were infected intranasally with 0.1 LD50 of influenza A/WSN/33 (WSN)-OVA one day later (an influenza strain engineered to express OVA257–264 in the neuraminidase stalk protein of influenza A/WSN/33). Recipient mice of OTII cells were infected intranasally with 500 VFU of PR8-OTII influenza virus (an engineered strain of PR8 that express the OVA323–339 peptide in the hemagglutinin (HA) molecule) one day after adoptive transfer. WSN-OVA and PR8-OTII were administered intranasally in 100 µl of PBS.
Chisolm D.A., Savic D., Moore A.J., Ballesteros-Tato A., León B., Crossman D.K., Murre C., Myers R.M, & Weinmann A.S. (2017). CCCTC-Binding Factor Translates Interleukin 2- and α-Ketoglutarate-Sensitive Metabolic Changes in T Cells into Context-Dependent Gene Programs. Immunity, 47(2), 251-267.e7.
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5

Isolation and Culture of CD4+ and CD8+ T-cells

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CD4+ and CD8+ T-cells were isolated from fresh and cryoconserved NAF, using anti-CD4 or anti-CD8 MACS-beads (Miltenyi) respectively, according to the manufacturer’s instructions. The yield was between 5–10% for CD8+ and 10–20% for CD4+ T-cells. The isolated T-cells were cultured overnight at a concentration of 1–2 × 106 cells/ml in T-cell medium consisting of RPMI 1640, 10% heat-inactivated human serum (Sigma-Aldrich), 2 mM L-glutamine, 20 mg/l gentamycin, 10 mM hydroxyethyl piperazineethanesulfonic acid (HEPES; PAA Labortechnik, Pasching, Austria), 1 mM sodium pyruvate (PAA), and 1% MEM non-essential amino acids (aa) (100×, PAA), supplemented with 20 U/ml IL-7 (Peprotech) for CD8+ T-cells and additionally with 5 ng/ml IL-15 (R&D systems, Wiesbaden-Nordenstadt, Germany) for CD4+ T-cells.
Gerer K.F., Erdmann M., Hadrup S.R., Lyngaa R., Martin L.M., Voll R.E., Schuler-Thurner B., Schuler G., Schaft N., Hoyer S, & Dörrie J. (2017). Preclinical evaluation of NF-κB-triggered dendritic cells expressing the viral oncogenic driver of Merkel cell carcinoma for therapeutic vaccination. Therapeutic Advances in Medical Oncology, 9(7), 451-464.
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6

T Cell Transfer for Tumor Immunotherapy

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For T cell transfer, T circulating CD8+ T cells from the spleen, or T resident memory cells from the skin inguinal adipose tissue of survivor mice were isolated using anti-CD8 MACS beads (Miltenyi Biotec). Enriched cells were then sorted as live CD45+CD3+CD8+ cells using FACS ARIA. Purity post-sorting was > 97%. Sorted T cells were then stimulated for 24 h in 24-well plates (~2 × 106 cells/well) coated with anti-CD3 (4 μg/ml) and anti-CD28 (4 μg/ml) in the presence of IL-2 (Peprotech, 50ng/ml). 5.×105 of activated splenic or skin inguinal adipose CD8+T cells were transferred by i.v. or intra dermal (i.d.) injection respectively on the right flank into naïve recipient mice 7 days prior tumor challenge. Some mice received both T cell subsets 7 days prior tumor challenge.
Zhivaki D., Borriello F., Chow O.A., Doran B., Fleming I., Theisen D.J., Pallis P., Shalek A.K., Sokol C.L., Zanoni I, & Kagan J.C. (2020). Inflammasomes within Hyperactive Murine Dendritic Cells Stimulate Long-Lived T Cell-Mediated Anti-tumor Immunity. Cell reports, 33(7), 108381.
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