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As a global market leader with numerous subsidiaries and distributors, Miltenyi Biotec is committed to providing our customers around the world with the highest quality products. In addition to direct selling in more than 20 countries in North America, Europe and Asia/Pacific, Miltenyi Biotec also provides support for our customers through an extensive distributor network covering dozens of additional countries.
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In this application protocol, we generate highly purified and viable microglia from adult mouse or rat brain tissue. Brain tissue from mice or rats older than P7 is dissociated into single-cell suspensions using the Adult Brain Dissociation Kit. The extracellular matrix is enzymatically digested using the kit components, while the gentleMACS™ Dissociator with Heaters is used for the mechanical dissociation steps during the on-instrument enzyme incubation. After the dissociation, the myelin and cell debris are removed using the Debris Removal Solution and is followed by subsequent removal of erythrocytes using the Red Blood Cell Removal Solution. The CD11b (Microglia) MicroBeads, mouse and human or the CD11b/c (Microglia) MicroBeads, rat are used to isolate microglia from the single-cell suspension.
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Enzyme mix 1 | Enzyme mix 2 | ||
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Enzyme P 50 µL | Buffer Z 1900 µL | Buffer Y 20 µL | Enzyme A 10 µL |
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Debris Removal Solution | D-PBS | Overlay (D-PBS) | |
---|---|---|---|
1 brain (400-500 mg) | 900 µL | 3100 µL | 4 mL |
2 brains (800-1000 mg) | 1800 µL | 6200 µL | 4 mL |
▲ Note: In case of very small amount of tissue (< 100 mg) cell debris removal can be performed in a 5 mL reagent tube using 450 µL of Debris Removal Solution, 1550 µL of D-PBS for resuspension of the cell pellet, and 2000 µL D-PBS for overlay.
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Unless otherwise specifically indicated, Miltenyi Biotec products and services are for research use only and not for therapeutic or diagnostic use.
autoMACS, CliniMACS, the CliniMACS logo, CliniMACS Prodigy, CryoMACS, CytoMix, CytoStim, DendriMACS, ExiTron, ExpAct, FeraSpin, FeraTrack, GadoSpin, gentleMACS, LIFE 18, LIFE 21, MACS, the MACS logo, MACSductin, MACSelect, MACSfectin, MACSflex, MACSiBead, MACSiMAG, MACSmix, MACSprep, MACSQuant, MACSQuantify, MACSxpress, MidiMACS, MiniMACS, miRXplore, MultiMACS, NeuroBrew, NiraWave, OctoMACS, PepTivator, pMACS, PolySon, PrepProtect, QuadroMACS, REAfinity, REAlease, Rheo, StemMACS, StraightFrom, SuperAmp, SuperMACS, TexMACS, TheraSorb, thermoMACS, TransAct, Tyto, the Tyto logo, VarioMACS, Vio, Viobility, VioBlue, VioBright, VioGreen, Viscover, and µMACS are registered trademarks or trademarks of Miltenyi Biotec B.V. & Co. KG and/or its affiliates in various countries worldwide. Ficoll-Paque is a trademark of GE Healthcare companies. Vectofusin-1 is a registered trademark of Genethon. All other trademarks mentioned in this document are the property of their respective owners and are used for identification purposes only. Copyright © 2019 Miltenyi Biotec B.V. & Co. KG and/or its affiliates. All rights reserved.
Column | Max. number of labeled cells | Max. number of total cells | Selector |
---|---|---|---|
MS | 1 x10⁷ | 2 x10⁷ | MiniMACS™, OctoMACS™, VarioMACS, SuperMACS II |
LD | 2 x10⁷ | 4 x10⁷ | MidiMACS™, QuadroMACS™, VarioMACS, SuperMACS II |
autoMACS | 5 x10⁷ | 1 x108 | autoMACS Pro |
Copyright © 2021 Miltenyi Biotec and/or its affiliates. All rights reserved.