Microglia cells from either newborn (P0-1, male or female) or adult (P40-55, male) C57BL/6J, NKCC1fl/fl (WT), or NKCC1fl/fl ΔCx3CR1 (KO) mice were isolated by magnetic separation using anti-CD11b microbeads (Miltenyi Biotec, Germany), with slight modification of the protocol described by Otxoa-de-Amezaga [85 (link)]. After transcardial perfusion with ice-cold phosphate-buffered saline (PBS), the brain tissues (cortices and hippocampi) were enzymatically dissociated with Neural Tissue Dissociation Kit-P (#130-092-628; Miltenyi Biotec). Myelin was removed by MACS Myelin Removal Beads II (#130-096-733, Miltenyi Biotec), then cells in a single-cell suspension were magnetically labeled with MACS CD11b microbeads (#130-093-634, Miltenyi Biotec) and were separated using MS columns (#130-042-201, Miltenyi Biotec, Germany). Cells selected with CD11b microbeads were plated onto poly-L-lysine precoated 96-well or 386-well plates at 3 × 104 cell/cm2 density and were cultured at 37°C in a 95% air/5% CO2 incubator in DMEM/Glutamax medium (#31966–021, Gibco) supplemented with 10% fetal bovine serum (FBS, #FB-1090, Biosera), 1% Pen/Strep (10,000 U/ml; #15140–122, ThermoFisher Scientific), and 10 nM macrophage colony-stimulating factor (M-CSF; #PMC2044, ThermoFisher Scientific) for 10 days.
Macs myelin removal beads 2
Manufactured by Miltenyi Biotec
Sourced in Germany
MACS Myelin Removal Beads II are magnetic microbeads designed for the depletion of myelin from single-cell suspensions. They enable the efficient removal of myelin, which can otherwise interfere with cell analysis and downstream applications.
Automatically generated - may contain errors
Lab products found in correlation
Pen strep, by Thermo Fisher Scientific (4 mentions)
Anti cd11b microbead, by Miltenyi Biotec (2 mentions)
Cd11b microbeads, by Thermo Fisher Scientific (2 mentions)
Macs cd11b microbead, by Miltenyi Biotec (2 mentions)
Neural tissue dissociation kit p, by Miltenyi Biotec (2 mentions)
Dmem glutamax medium, by Thermo Fisher Scientific (2 mentions)
Ms column, by Miltenyi Biotec (2 mentions)
Fetal bovine serum (fbs), by Biosera (2 mentions)
M csf, by Thermo Fisher Scientific (1 mentions)
2 protocols using macs myelin removal beads 2
1
Isolation and culture of primary murine microglia
2
Isolation and Culture of Mouse Microglia
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Microglia cells from either newborn (P0-1, male or female) or adult (P40-55, male) C57BL/6J, NKCC1 fl/fl (WT) or NKCC1 fl/fl Δ Cx3CR1 (KO) mice were isolated by magnetic separation using anti-CD11b microbeads (Miltenyi Biotec, Germany), with slight modification of the protocol described by Otxoa-de-Amezaga [81] (link). After transcardial perfusion with ice-cold PBS (phosphate buffered saline), the brain tissues (cortices and hippocampi) were enzymatically dissociated with Neural Tissue Dissociation Kit-P (#130-092-628; Miltenyi Biotec). Myelin was removed by MACS Myelin Removal Beads II (#130-096-733, Miltenyi Biotec), then cells in a single-cell suspension were magnetically labeled with MACS CD11b microbeads (#130-093-634, Miltenyi Biotec) and were separated using MS columns (#130-042-201, Miltenyi Biotec, Germany). Cells selected with CD11b microbeads were plated onto poly-L-lysine precoated 96-well or 386-well plates at 3x10 4 cell/cm 2 density and were cultured at 37 °C in a 95% air/5% CO2 incubator in DMEM/Glutamax medium (#31966-021, Gibco) supplemented with 10% FBS (fetal bovine serum, #FB-1090, Biosera), 1% Pen/Strep (10000 U/ml; #15140-122, ThermoFisher Scientific) and 10 nM Macrophage Colony-Stimulating Factor (M-CSF; #PMC2044, ThermoFisher Scientific) for 10 days.
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