Macs separation ms columns

Manufactured by Miltenyi Biotec

MACS separation MS columns are designed for magnetic cell separation. They provide a platform for the isolation and enrichment of target cells from heterogeneous cell populations. The columns leverage the power of magnetic beads coated with specific antibodies to selectively capture and separate the desired cell types.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Crpmi, by Thermo Fisher Scientific (1 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) Histopaque 1077, by Merck Group (1 mentions) Cd14 microbead, by Miltenyi Biotec (1 mentions) Cd11b magnetic microbeads, by Miltenyi Biotec (1 mentions) Adult brain dissociation kit, by Miltenyi Biotec (1 mentions) Anti fitc microbeads, by Miltenyi Biotec (1 mentions) Perfect count microspheres cyt pcm 100, by Cytognos (1 mentions) Lineage cell depletion kit, by Miltenyi Biotec (1 mentions) Facsdiva, by BD (1 mentions) Sytox green, by Thermo Fisher Scientific (1 mentions) Facscanto 2, by BD (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Methocult gf m3534, by STEMCELL (1 mentions)

5 protocols using macs separation ms columns

Isolation and Culture of PBMCs

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Blood from donors was collected and processed within 2 h. PBMCs were separated using Histopaque 1077 at room temperature (Sigma-Aldrich) and following the protocol for mononuclear cell separation of SepMate-50 tubes (StemCells). Cells were resuspended in complete RPMI-1640 medium (10% Fetal bovine serum, 1% sodium pyruvate, 1% HEPES and 1% l-glutamine; cRPMI) (GIBCO), counted and plated in 24-well plates at a concentration of 0.5 × 10⁶ cell mL^–1 in 500 μl of cRPMI.
CD14 + cells were isolated from cryopreserved PBMCs. PBMCs were magnetic labeled with CD14 MicroBeads (Miltenyi Biotec) and isolated through positive selection using MACS Separation Columns MS (Miltenyi Biotec), according to the manufacturer’s protocol. Purified cells (~95% pure) were counted and plated in 96 well plates at a concentration of 0.5 × 10⁶ cell mL^–1 in 200 μl of cRPMI (GIBCO).

Sousa J., Cá B., Maceiras A.R., Simões-Costa L., Fonseca K.L., Fernandes A.I., Ramos A., Carvalho T., Barros L., Magalhães C., Chiner-Oms Á., Machado H., Veiga M.I., Singh A., Pereira R., Amorim A., Vieira J., Vieira C.P., Bhatt A., Rodrigues F., Rodrigues P.N., Gagneux S., Castro A.G., Guimarães J.T., Bastos H.N., Osório N.S., Comas I, & Saraiva M. (2020). Mycobacterium tuberculosis associated with severe tuberculosis evades cytosolic surveillance systems and modulates IL-1β production. Nature Communications, 11, 1949.

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Isolation of Murine Microglia

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Mice were anesthetized and perfused intracardially with ice-cold Dulbecco’s phosphate-buffered saline to harvest brains. After the subsequent tissue dissociation, debris removal and red blood cell removal procedures, Adult Brain Dissociation Kit (130-107-677, Miltenyi Biotec) was used to generate single-cell suspension. Microglia were further isolated from the single-cell suspension using MACS Separation Columns (MS) (130-042-201, Miltenyi Biotec) and magnetic CD11b Microbeads (130-093-634, Miltenyi Biotec).

Ding X., Wang J., Huang M., Chen Z., Liu J., Zhang Q., Zhang C., Xiang Y., Zen K, & Li L. (2021). Loss of microglial SIRPα promotes synaptic pruning in preclinical models of neurodegeneration. Nature Communications, 12, 2030.

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Enrichment of Ductal Cells via DBA Lectin Sorting

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To enrich extracted cell populations for ductal cell extracts, DBA Lectin sorting was completed. Cells were isolated from a mouse pancreata as described above and suspended with 1:200 diluted DBA lectin–FITC (Vector Labs, cat. no. FL-1031) and incubated for 10 min in the dark at 4 °C on a rotor. Cells were washed with 700 μl of separation buffer (PBS, 0.5% BSA, and 2mM EDTA) and centrifuged for 10 min at 300g at 4 °C. The pellet was resuspended in 90 μl of separation buffer and 10 μl of anti-FITC MicroBeads (Miltenyi Biotec, cat. no. 130-048-701) and incubated on a rotor at 4 °C for 15 min. The cells were washed with 1mL of sorting buffer (PBS 0.5% BSA, 2mM EDTA) and the pellet (300g for 10 min) was resuspended in 500 μl of sorting buffer. MACS Separation (MS) columns (Miltenyi Biotec, cat. no. 130-042-201) were prepared by running 500 μl of chilled sorting buffer and then the labeled cell suspensions (500 μl) were applied to the column. The columns were washed three times with 500 μl of buffer and the flow through was collected as the DBA lectin–negative fraction. The column was removed from the magnetic field and 1,000 μl of separation buffer was then added with a plunger to collect the DBA lectin–positive fraction. The samples were spun down (300g for 15 min at 4 °C) and the pellets used for downstream applications.

Das K.K., Heeg S., Pitarresi J.R., Reichert M., Bakir B., Takano S., Kopp J.L., Wahl-Feuerstein A., Hicks P., Sander M, & Rustgi A.K. (2018). ETV5 regulates ductal morphogenesis with Sox9 and is critical for regeneration from pancreatitis. Developmental dynamics : an official publication of the American Association of Anatomists, 247(6), 854-866.

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Comprehensive Immunophenotyping of Murine Hematopoietic Cells

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Blood, BM cells and splenocytes were collected as previously described^{19 (link),40 (link)}. The different cell samples (0.5–2 × 10⁶ cells) were stained for the indicated surface markers (see Supplemental Table S2 online) for 20 min at room temperature and subsequently washed twice with buffer A. We distinguished live and dead cells by adding SYTOX Green (S7020, Life Technologies) or DAPI (D9542, Sigma-Aldrich) 5 min before FACS analysis. Haematopoietic progenitors were isolated from the spleen and BM by magnetic-activated cell sorting (MACS) using a Lineage Cell Depletion Kit (130-090-858, Miltenyi) and MACS separation MS columns (130-042-201, Miltenyi) prior staining. Unstained cells were used as a negative control to establish the flow cytometer voltage settings, and single-colour positive controls were used to adjust compensation. The absolute number of cells was calculated by adding Perfect-Count Microspheres (CYT-PCM-100, Cytognos) to the flow cytometry samples. Apoptosis, cell cycle, and cell proliferation were determined by flow cytometry as previously described^{19 (link),40 (link)}. The flow cytometry data were acquired using a FACSCanto II and analysed with FACSDiva (Becton and Dickinson) or FlowJo software.

Sánchez Á., Orizaola M.C., Rodríguez-Muñoz D., Aranda A., Castrillo A, & Alemany S. (2020). Stress erythropoiesis in atherogenic mice. Scientific Reports, 10, 18469.

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Isolation and Culture of Hematopoietic Progenitors

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Haematopoietic progenitors were isolated from the BM of DsRed⁺Wt and Map3k8^−/− mice by magnetic-activated cell sorting (MACS) using a LIN⁻ lineage depletion kit (containing antibodies against CD5, CD11b, GR-1, 7-4, and Ter119) and MACS separation MS columns (Miltenyi). Isolated LIN⁻ DsRed⁺Wt and Map3k8^−/− cells (2,500) were plated at a 1:1 ratio in a well with methylcellulose medium, which contains insulin, H transferrin, stem cell factor, IL-3, and IL-6 (MethoCult GF M3534, StemCell Technologies) in the presence or absence of LPS (500 ng/ml). After six days of incubation at 37 °C in an atmosphere containing 5% CO₂, the cells were harvested and subjected to FACS analysis with Perfect-Count microspheres.

Sánchez Á., Relaño C., Carrasco A., Contreras-Jurado C., Martín-Duce A., Aranda A, & Alemany S. (2017). Map3k8 controls granulocyte colony-stimulating factor production and neutrophil precursor proliferation in lipopolysaccharide-induced emergency granulopoiesis. Scientific Reports, 7, 5010.

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