Anti cd8 magnetic beads

Manufactured by Miltenyi Biotec

Anti-CD8 magnetic beads are a laboratory product designed for the isolation and enrichment of CD8+ T cells from biological samples. The beads are coated with antibodies specific to the CD8 surface antigen, allowing for the separation and concentration of CD8-positive cells using a magnetic field. This core function enables researchers to study and analyze CD8+ T cell populations, which play a crucial role in immune responses and various disease processes.

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Lab products found in correlation

Uea 1, by Merck Group (1 mentions) M5 114, by Thermo Fisher Scientific (1 mentions) Facscanto flow cytometer, by BD (1 mentions) Anti cd4, by Miltenyi Biotec (1 mentions) α mhcii, by Thermo Fisher Scientific (1 mentions) αcd45 30 f11, by BioLegend (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) β mercaptoethanol, by Thermo Fisher Scientific (1 mentions) Rbc lysis buffer, by Merck Group (1 mentions) Fty720, by Novartis (1 mentions) Ls column, by Miltenyi Biotec (1 mentions) Anti cd16 32, by BioXCell (1 mentions) Anti cd11c magnetic beads, by Miltenyi Biotec (1 mentions) Celesta, by BD (1 mentions) Intracellular staining permeabilization wash buffer, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Magnetic beads (547 citations) CD8 magnetic beads (10 citations) Anti-CD8 (8 citations) Anti-CD8 beads (5 citations) Magnetic bead-conjugated anti-mouse CD8 (53–6.7) monoclonal antibody (2 citations)

6 protocols using anti cd8 magnetic beads

Isolation and Characterization of Immune Cells

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Human BLS cell lines and in vitro generated B cell mutants have been described and are established human cell lines [1 (link),30 (link)]. T cells were enriched using anti-CD4 and/or anti-CD8 magnetic beads (Miltenyi Biotec). For all flow cytometric analyses, gating on living cells and exclusion of doublets was performed. Enriched TEC suspensions [2 (link)] were washed in PBS, 2% FCS, 5mM EDTA and stained for flow cytometry using death exclusion markers (either DAPI or 7AAD), UEA1 (Sigma) and the following mAb-conjugated mix: α-CD45 (30F11, BioLegend) and α-BP1 (6C3; BioLegend), α-MHCII (M5/114.15.2; eBioscience) and α-EpCAM (G8.8, Developmental Studies Hybridoma Bank, Iowa), α-H2-Db (B22/24g) and α-H2-Kb (B8.24.3). Splenocytes were preincubated with anti-CD16/32 (2.4G2) to block FcRs and stained using Abs against CD8a (Ly-2), CD3e (145-2C11), CD4 (L3T4), CD11b (M1/70), CD11c (N418), CD19 (1D3), H2-Db (28-14-8), H2-Kb (AF6–88.5.5.3), MHCII (M5/114.15.2), NK1.1 (PK136), B220 (RA3-6B2), Qa-2 (69H1-9-9) (all from eBioscience). Streptavidin conjugated to different fluorophores was from eBioscience. Stainings were performed with appropriate combinations of fluorophores. Data was acquired with a FACSCanto flow cytometer (Becton Dickinson) and analyzed using FlowJo software (Tree Star).

Ludigs K., Seguín-Estévez Q., Lemeille S., Ferrero I., Rota G., Chelbi S., Mattmann C., MacDonald H.R., Reith W, & Guarda G. (2015). NLRC5 Exclusively Transactivates MHC Class I and Related Genes through a Distinctive SXY Module. PLoS Genetics, 11(3), e1005088.

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Evaluating T-cell Cytotoxicity against Tumor Cells

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C57BL/6 mice were injected with 10⁶ EL4 cells, and when tumors reached 50 mm³, mice were treated with Val-ILs. At day 12, T8 lymphocytes were recovered from tumors and spleens, with anti-CD8 magnetic beads (130-104-075, Miltenyi Biotec) and cultured in RPMI-1640 media supplemented with 10% FBS, 1% PSA, MEM 1× (11140035, Thermo Fisher Scientific), l-glutamine 2 mM (25030149, Thermo Fisher Scientific), HEPES 10 mM (15630080, Thermo Fisher Scientific), pyruvate sodium 1 mM (11360070, Thermo Fisher Scientific) and β-mercaptoethanol 5.5 nM (21985023, Thermo Fisher Scientific). T8 lymphocytes were enumerated and co-cultured with EL4 cells, at a ratio of 1:100. Viability was assessed by XTT cell proliferation assay (Thermo Fisher Scientific), following 48 h of treatment.

Georgievski A., Bellaye P.S., Tournier B., Choubley H., Pais de Barros J.P., Herbst M., Béduneau A., Callier P., Collin B., Végran F., Ballerini P., Garrido C, & Quéré R. (2024). Valrubicin-loaded immunoliposomes for specific vesicle-mediated cell death in the treatment of hematological cancers. Cell Death & Disease, 15(5), 328.

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Adoptive Transfer of Activated CD8+ T Cells

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Bulk H-2K^b+, ovalbumin-specific OT-I Thy1.1⁺ T cells were adoptively transferred into B6 mice, which were then immunized by SC, IV, or IP routes with SIINFEKL peptide-pulsed, activated bone marrow–derived dendritic cells (BMDCs) (9 (link)). Alternatively, CD8⁺ T cells from FH or FH × CXCR3^−/− transgenic mice were adoptively transferred into AAD mice, and immunized with YMDGTMSQV peptide-pulsed activated BMDC. FTY720 (Novartis) was administered daily to retain effector CD8⁺ T cells in draining lymph nodes (LNs). On day 5, draining LNs and/or spleens were homogenized and treated with RBC lysis buffer (Sigma). CD8⁺ T cells were enriched using anti-CD8⁺ magnetic beads (Miltenyi) and 300,000-500,000 were injected IV into tumor-bearing animals.

Woods A.N., Wilson A.L., Srivinisan N., Zeng J., Dutta A.B., Peske J.D., Tewalt E.F., Gregg R.K., Ferguson A.R, & Engelhard V.H. (2017). Differential expression of homing receptor ligands on tumor associated vasculature that control CD8 effector T cell entry. Cancer immunology research, 5(12), 1062-1073.

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Adoptive Transfer of OT I Cells for Lymphoma

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Three groups of female 5 weeks old C57Bl/6 CD45.1 mice, five animals per group, were injected subcutaneously with OVA expressing CD45+ E. G7-OVA lymphoma tumor cells. Seven days post-tumor cell injection, when tumor volume reached ~100 mm³, two groups of mice were treated with 3.7 MBq ¹³¹I-30F11 (20 μg), while the third group was left untreated. Four days post-lymphodepletion, OVA-specific CD8+ T cells were purified using anti-CD8 magnetic beads from CD45.2 OT I mice according to the manufacturer’s protocol (Miltenyi Biotec, Auburn, CA). One group of mice treated with 3.7 MBq ¹³¹I-30F11 was given a single i. v. injection of 10⁶ CD45.2 OT I CD8+ cells. Tumor volume and body weight were monitored, and mice were sacrificed when tumor volume exceeded 4,000 mm³ or tumors became necrotic. Blood and spleen were assessed for immune cell subsets and presence of engrafted CD45.2 OT I cells by flow cytometry.

Dawicki W., Allen K.J., Garg R., Geoghegan E.M., Berger M.S., Ludwig D.L, & Dadachova E. (2020). Targeted lymphodepletion with a CD45-directed antibody radioconjugate as a novel conditioning regimen prior to adoptive cell therapy. Oncotarget, 11(39), 3571-3581.

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Purification and Labeling of OTI CD8+ T Cells

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OTI cells were purified from the spleens of CD45.1+ OTI mice by incubating single cell suspensions with anti-CD16/32 (BioXcell) and anti–CD8 magnetic beads (Miltenyi Biotec), for 45 minutes at 4°C. After washing, CD8⁺ cells were enriched by magnetic separation using LS–columns (Miltenyi Biotec) and stained with cell trace violet (CTV, Thermo–Fisher) according to instructions. Cells were adjusted to 6 x 10⁵ cells/mL in sterile PBS and 50 μL of the cell suspension were transferred intraperitoneally. For DC enrichment, cell suspensions from dissociated lungs were incubated with anti–CD11c magnetic beads followed by magnetic separation using LS columns (Miltenyi Biotec). After enrichment, DCs were purified by first gating on Lin⁻CD64⁻MHCII⁺CD11c⁺ cells and sorted based on expression of CD103, CD11b, SIRPα. Post–sorting purity was >95%. All sorting was performed using a FACSAriaII provided by the Flow Cytometry and Single Cell services Core at UAB.

, & Nagylaki T. (1991). Error bounds for the fundamental and secondary theorems of natural selection.. Proceedings of the National Academy of Sciences of the United States of America, 88(6), 2402-2406.

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Immune Cell Profiling in Stressed Rats

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Lymph nodes or blood samples were subjected to flow cytometry using Celesta (BD) and the following fluorescence-labeled antibodies: BV510-conjugated anti-CD45, APC-conjugated anti-CD3, FITC-conjugated anti-CD4, Percp-5.5-conjugated anti-CD8, and PE-conjugated anti-CD45RC. CD8+T cells were isolated from the lympho-nodes (LNs) of adult rats using anti-CD8 magnetic beads (Miltenyi Biotec). For intracellular cytokine staining, T cells were obtained immediately from the CLNs of stressed rats and then permeabilized using intracellular staining permeabilization wash buffer (eBioscience, 88-8824-00). Then, intracellular CCL5 was analyzed by flow cytometry.

Shi D.D., Zhang Y.D., Zhang S., Liao B.B., Chu M.Y., Su S., Zhuo K., Hu H., Zhang C, & Wang Z. (2023). Stress-induced red nucleus attenuation induces anxiety-like behavior and lymph node CCL5 secretion. Nature Communications, 14, 6923.

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