Automacs separation system

Manufactured by Miltenyi Biotec

Sourced in Germany

The AutoMACS separation system is a fully automated magnetic cell separation instrument designed for efficient and reproducible cell isolation. It utilizes magnetic microbeads conjugated to specific antibodies to selectively label target cells, which are then separated using a magnetic field. The system allows for automated processing of multiple samples with high purity and recovery rates.

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Lab products found in correlation

Aria 2, by BD (1 mentions) Macs cd4 t cell isolation kit, by Miltenyi Biotec (1 mentions) Facsaria cell sorter, by BD (1 mentions) Anti cd14 magnetic microbeads, by Miltenyi Biotec (1 mentions) Dulbecco s modified eagle s medium, by Fujifilm (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Hi fbs, by Thermo Fisher Scientific (1 mentions) Phorbol 12 myristate 13 acetate, by Merck Group (1 mentions) Optiprep, by Axis-Shield (1 mentions) Biotinylated anti cd3 clone 145 2c11, by BD (1 mentions) Biotinylated anti cd28 clone 37, by BD (1 mentions) Penicillin, by Merck Group (1 mentions) Streptomycin, by Merck Group (1 mentions) Ionomycin, by Merck Group (1 mentions) Facsaria 2, by BD (1 mentions)

Spelling variants of this product

AutoMACS (460 citations) AutoMACS system (69 citations) AutoMacs magnetic separation system (8 citations) AutoMACS Pro Separation System (7 citations) AutoMACS separation (4 citations) AutoMACS magnetic cell separation system (4 citations) AutoMACS cell separation system (2 citations)

6 protocols using automacs separation system

Isolation and Sorting of Murine T Cell Subsets

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For the in vitro assays, peripheral CD4⁺ T cells were enriched from pooled spleen and lymph node cells from Foxp3^RFP reporter mice (C57BL/6) using direct anti-CD4 (L3T4) microbeads followed by magnetic separation using the autoMACS separation system (Miltenyi Biotec). Subsequently, enriched cells were labeled with respective antibodies and sorted as CD4⁺CD90.2⁺Foxp3^RFP− peripheral T cells on ARIA II (BD Biosciences) or MoFlo (Beckman Coulter). For the in vivo assay, naïve CD4⁺ T cells were isolated from pooled spleen and lymph node cells from Foxp3^hCD2 CD45.1 reporter mice. Briefly, CD4⁺ T cells were enriched as described above and labeled with respective antibodies. Subsequently, naïve CD4⁺ T cells (CD4⁺CD90.2⁺CD45.1⁺CD25⁻CD44⁻CD62L^hiFoxp3^hCD2−) were sorted on ARIA II or MoFlo.

Nikolouli E., Hardtke-Wolenski M., Hapke M., Beckstette M., Geffers R., Floess S., Jaeckel E, & Huehn J. (2017). Alloantigen-Induced Regulatory T Cells Generated in Presence of Vitamin C Display Enhanced Stability of Foxp3 Expression and Promote Skin Allograft Acceptance. Frontiers in Immunology, 8, 748.

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Adoptive Transfer of Murine CD4+ T Cells

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CD4⁺ T cells were isolated from spleens of 5- to 8-wk-old female C57BL/10SgSnAi mice via negative selection using the MACS CD4 T cell isolation kit (Miltenyi Biotec Inc, Auburn, CA) and the AutoMACS separation system (Miltenyi Biotec). Enriched cells were subsequently stained for CD4⁺ and CD45RB and sorted by flow cytometry into fractions containing the 15% of CD4⁺ cells with the brightest CD45RB-staining (CD4⁺CD45RB^high) and the 35% with the lowest CD45RB-staining (CD4⁺CD45RB^low) cells using a FACS Aria cell sorter (BD Biosciences). Sorted cells were >95% pure. A total of 2 × 10⁵ CD45RB^high cells or 2 × 10⁵ CD45RB^high plus 2 × 10⁵ CD45RB^low were transferred i.p. to recipient 5- to 8-wk-old female RAG-2^−/− mice. Non-transferred RAG-2^−/− mice served as controls. Mice were followed for the development of inflammatory changes of eyelids, and haziness of the cornea, which correlates with histological evidence of keratitis.

Seo K.Y., Kitamura K., Han S.J, & Kelsall B. (2017). Th17 cells mediate inflammation in a novel model of spontaneous experimental autoimmune lacrimal keratoconjunctivitis with neural damage. The Journal of allergy and clinical immunology, 142(1), 96-108.e2.

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Isolating Primary Myeloma Cells

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Two BM samples from untreated myeloma patients at diagnosis were used to isolate primary myeloma cells via CD138-positive selection using the AutoMACs separation system (Miltenyi-Biotec) with a final purity > 95%. These primary myeloma cells were subsequently used in co-culture studies with pMSCs.

Garcia-Gomez A., Las Rivas J.D., Ocio E.M., Díaz-Rodríguez E., Montero J.C., Martín M., Blanco J.F., Sanchez-Guijo F.M., Pandiella A., San Miguel J.F, & Garayoa M. (2014). Transcriptomic profile induced in bone marrow mesenchymal stromal cells after interaction with multiple myeloma cells: implications in myeloma progression and myeloma bone disease. Oncotarget, 5(18), 8284-8305.

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Differentiation of THP1 and CD14+ Monocytes

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THP1 cells, a human acute monocytic cell line that was purchased from the Human Science Foundation (Tokyo, Japan), were cultured in RPMI-1640 supplemented with 10% HI-FBS (Thermo Fisher Scientific, Waltham, MA, USA). Differentiation was achieved by re-suspending the cells at 8 × 10⁵ cells/ml in Dulbecco’s modified Eagle’s medium (Wako) supplemented with 10% heat-inactivated fetal bovine serum and penicillin-streptomycin with the addition of 100 nM phorbol 12-myristate 13-acetate (Sigma) for 48 hours. Non-adherent cells were removed by aspiration of the supernatant followed by replacement with fresh medium.
The human blood PBMC was obtained by using Optiprep (Axis-Shield, Dundee, Scotland) in accordance with the manufacturer’s instructions. CD14⁺ monocytes were then isolated from PBMC by the AutoMacs separation system using anti-CD14 magnetic microbeads (Miltenyi Biotec, Germany). The CD14⁺ human monocytes were differentiated to macrophages by commercially available macrophage differentiation kit (R&D systems, Minnesota).

Inoue M., Niki M., Ozeki Y., Nagi S., Chadeka E.A., Yamaguchi T., Osada-Oka M., Ono K., Oda T., Mwende F., Kaneko Y., Matsumoto M., Kaneko S., Ichinose Y., Njenga S.M., Hamano S, & Matsumoto S. (2018). High-density lipoprotein suppresses tumor necrosis factor alpha production by mycobacteria-infected human macrophages. Scientific Reports, 8, 6736.

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Isolation and Stimulation of CD4+ T Cells

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Total CD4⁺ T cells were isolated from spleen and LNs of CalDAG GEFI^–/–, CalDAG GEFI^+/– and CalDAG GEFI^+/+ mice using mouse CD4 DirectBeads (L3T4) and the autoMACS separation system (Miltenyi Biotec). For stimulation, 1 × 10⁶ cells were resuspended in complete RPMI (cRPMI; 10% FCS, 25,000 Units penicillin/25 mg streptomycin, 1 mM sodium pyruvate, 25 mM HEPES, 50 µM β-mercaptoethanol) supplemented with either 20 ng/ml PMA and 1 µg/ml ionomycin (both from Sigma) or 35 µM sodium pervanadate (Sigma), and incubated for 5 min at 37 °C. For anti-CD3/CD28 stimulation, cells were coated with 20 µg/ml biotinylated anti-CD3 (clone 145-2C11, BD Biosciences) and 10 µg/ml biotinylated anti-CD28 (clone 37.51, BD Biosciences), and crosslinking was induced for 5 min by addition of 10 µg/ml streptavidin in cRPMI. As stimulation controls, cells were either left on ice or incubated at 37 °C for 5 min in cRPMI. Following stimulation, cells were washed once in ice-cold PBS, and cell pellets were further processed for lysis and Western Blotting.

Niemz J., Kliche S., Pils M.C., Morrison E., Manns A., Freund C., Crittenden J.R., Graybiel A.M., Galla M., Jänsch L, & Huehn J. (2017). The Guanine-Nucleotide Exchange Factor Caldag Gefi Fine-Tunes Functional Properties of Regulatory T Cells. European Journal of Microbiology & Immunology, 7(2), 112-126.

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Isolation of Murine CD4+ T Cell Subsets

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For purification of CD4SP thymocytes from male or female donor mice, total thymocytes were depleted of CD8⁺ cells using APC-conjugated anti-CD8⁺ and anti-APC microbeads (Miltenyi Biotec) followed by magnetic separation using the autoMACS separation system (Miltenyi Biotec). Peripheral CD4⁺ T cells were enriched from pooled spleen and lymph node cells using anti-CD4 microbeads followed by magnetic separation using the autoMACS separation system. CD4SP Foxp3⁻ thymocytes or CD4⁺Foxp3⁻ peripheral T cells were sorted as CD4⁺RFP⁻GFP⁻ cells from Foxp3^RFP x IL-10^GFP reporter mice or stained with anti-human CD2 to isolate CD4SP Foxp3^hCD2− thymocytes from Foxp3^hCD2 mice on FACS Aria II (BD Biosciences). For the in vitro and in vivo suppression assays, naïve CD4⁺ T cells were isolated from pooled spleen and lymph node cells from CD45.1 congenic C57BL/6 mice. Briefly, CD4⁺ T cells were enriched using anti-CD4 microbeads followed by magnetic separation using the autoMACS separation system. Subsequently, naïve CD4⁺ T cells (CD4⁺CD90.2⁺CD25⁻CD44⁻CD62L^hi) were sorted on FACS Aria II.

Garg G., Nikolouli E., Hardtke-Wolenski M., Toker A., Ohkura N., Beckstette M., Miyao T., Geffers R., Floess S., Gerdes N., Lutgens E., Osterloh A., Hori S., Sakaguchi S., Jaeckel E, & Huehn J. (2017). Unique properties of thymic antigen-presenting cells promote epigenetic imprinting of alloantigen-specific regulatory T cells. Oncotarget, 8(22), 35542-35557.

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