MDSCs were generated as previously reported (35 (link)). Briefly, bone marrow nucleated cells were cultured in DMEM supplied with 2mM L-glutamine, 1.5g/L sodium bicarbonate, 10mM HEPES, 1.0mM sodium pyruvate (all from Gibco, MA, USA), 10% fetal bovine serum (Hyclone, MA, USA), 40ng/ml rmGM-CSF and 40ng/ml rmIL-6 (BioLegend), with or without 50% E.G7-OVA supernatant. The floating cells were collected on day 4 and enriched by anti-mouse Gr-1 Microbeads (Miltenyi). The cells with purity > 90% were used. In some experiments, the cells were stained with anti-mouse Ly6C, anti-mouse Ly6G, and anti-mouse/human CD11b (Biolegend) and sorted with BD FACS Aria II cell sorter (BD).
Anti mouse gr 1 microbeads
Manufactured by Miltenyi Biotec
Sourced in Germany
Anti-mouse Gr-1 Microbeads are a cell separation tool used for the isolation of Gr-1 positive cells from mouse samples. They consist of superparamagnetic beads coated with antibodies specific to the mouse Gr-1 antigen.
Automatically generated - may contain errors
Lab products found in correlation
Facsaria 2 cell sorter, by BD (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Hepes, by Thermo Fisher Scientific (1 mentions)
Rmgm csf, by BioLegend (1 mentions)
Rmil 6, by BioLegend (1 mentions)
Anti mouse ly6c, by BioLegend (1 mentions)
Anti mouse ly6g, by BioLegend (1 mentions)
2 protocols using anti mouse gr 1 microbeads
1
Isolation and Characterization of MDSCs
2
MDSC Adoptive Transfer in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
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MDSCs were isolated from the spleens of WT and Camkk2-/- mice 21 days after E.G7-OVA injection and were enriched by anti-mouse Gr-1 Microbeads (Miltenyi, Bergisch Gladbach, Germany) according to the manufacturer’s instructions. Then, 2×106 MDSCs (> 90% pure) were injected into Camkk2-/- mice through the tail vein, followed by 1×105 E.G7-OVA tumor cells injected subcutaneously into the flank. Tumor size was measured as described above.
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