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Cd11b conjugated magnetic microbeads

Manufactured by Miltenyi Biotec
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CD11b-conjugated magnetic microbeads are a type of lab equipment used for cell separation and isolation. They are composed of magnetic particles coated with antibodies that specifically bind to the CD11b antigen expressed on the surface of certain cell types. These beads can be used to isolate and enrich CD11b-positive cells from complex samples, such as blood or tissue, using magnetic separation techniques.

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Lab products found in correlation

Rlt buffer, by Qiagen (2 mentions) Rneasy mini kit, by Qiagen (2 mentions) Dnase 1, by Roche (2 mentions) Collagenase type 1, by Worthington (2 mentions) Percoll, by Merck Group (1 mentions) Ls column, by Miltenyi Biotec (1 mentions)

3 protocols using cd11b conjugated magnetic microbeads

1

Isolation and Purification of Adult Microglia

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Adult microglia were isolated based on a modified version of an established protocol89 (link) LH rats were deeply anaesthetised with sodium pentobarbital (Euthatal®, 80 mg/kg, administered intraperitoneally) and transcardially perfused with ice-cold DPBS containing 2 mM EDTA. Brains were dissected, homogenised with Dounce homogeniser in 5 ml ice-cold DPBS, filtered through a 70 µm cell strainer to remove any debris and centrifuged (300 × g, 7 min, 4 °C). Pellets were resuspended in 7 ml of 30% Percoll® (Sigma, P1644) and centrifuged (800×g, 30 min, room temperature), to remove myelin. Microglia were isolated from the pellet with CD11b conjugated magnetic microbeads (Miltenyi Biotec) according to manufacturer’s protocol. Magnetic-activated cell sorted CD11b+ cells were flushed from LS columns (Miltenyi Biotech) and collected in complete medium DMEM/F12 supplemented with 10% FBS and Pen-Strep. Cells were seeded at 100,000 cells/cm2 concentration in 24-well plates coated with poly-d-lysine (0.1 mg/ml. Sigma-Aldrich).
Francos-Quijorna I., Sánchez-Petidier M., Burnside E.R., Badea S.R., Torres-Espin A., Marshall L., de Winter F., Verhaagen J., Moreno-Manzano V, & Bradbury E.J. (2022). Chondroitin sulfate proteoglycans prevent immune cell phenotypic conversion and inflammation resolution via TLR4 in rodent models of spinal cord injury. Nature Communications, 13, 2933.
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2

Isolation of Microglia from Human Brain

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Brain tissue was provided by the Netherlands Brain Bank (NBB). Informed consent for brain autopsy, the use of tissue, and use of clinical information was obtained pre-mortem. The procedures of the NBB are in accordance with all national laws and the procedures have been approved by the ethics committee of the VU University Medical Center (Amsterdam, Netherlands). Brain tissues were collected from one male and three female donors without a history of a neurological or psychiatric disorder with an age range between 60 and 77 years. Microglia were isolated from the medial frontal gyrus and temporal superior gyrus as described before64 . In short, 2–10 g of tissue was collected within 13 hours after death and stored in Hibernate medium on ice, and the isolation procedure was started within 2 to 24 h after autopsy. A single-cell suspension was generated by dissociating the tissue mechanically through a metal sieve, followed by an enzymatic digestion using collagenase Type I (3,700 U/ml; Worthington) and DNaseI (200 μg/ml; Roche). Microglia were further purified using a Percoll gradient and positive selection using CD11b-conjugated magnetic microbeads (Miltenyi Biotec). Microglia were lysed in RLT buffer (Qiagen), and RNA was extracted according to the protocol provided by the RNeasy Mini Kit (Qiagen).
Guttikonda S.R., Sikkema L., Tchieu J., Saurat N., Walsh R., Harschnitz O., Ciceri G., Sneeboer M., Mazutis L., Setty M., Zumbo P., Betel D., de Witte L.D., Pe’er D, & Studer L. (2021). Fully defined human pluripotent stem cell-derived microglia and tri-culture system model C3 production in Alzheimer’s disease. Nature neuroscience, 24(3), 343-354.
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3

Isolation of Microglia from Human Brain

Check if the same lab product or an alternative is used in the 5 most similar protocols
Brain tissue was provided by the Netherlands Brain Bank (NBB). Informed consent for brain autopsy, the use of tissue, and use of clinical information was obtained pre-mortem. The procedures of the NBB are in accordance with all national laws and the procedures have been approved by the ethics committee of the VU University Medical Center (Amsterdam, Netherlands). Brain tissues were collected from one male and three female donors without a history of a neurological or psychiatric disorder with an age range between 60 and 77 years. Microglia were isolated from the medial frontal gyrus and temporal superior gyrus as described before64 . In short, 2–10 g of tissue was collected within 13 hours after death and stored in Hibernate medium on ice, and the isolation procedure was started within 2 to 24 h after autopsy. A single-cell suspension was generated by dissociating the tissue mechanically through a metal sieve, followed by an enzymatic digestion using collagenase Type I (3,700 U/ml; Worthington) and DNaseI (200 μg/ml; Roche). Microglia were further purified using a Percoll gradient and positive selection using CD11b-conjugated magnetic microbeads (Miltenyi Biotec). Microglia were lysed in RLT buffer (Qiagen), and RNA was extracted according to the protocol provided by the RNeasy Mini Kit (Qiagen).
Guttikonda S.R., Sikkema L., Tchieu J., Saurat N., Walsh R., Harschnitz O., Ciceri G., Sneeboer M., Mazutis L., Setty M., Zumbo P., Betel D., de Witte L.D., Pe’er D, & Studer L. (2021). Fully defined human pluripotent stem cell-derived microglia and tri-culture system model C3 production in Alzheimer’s disease. Nature neuroscience, 24(3), 343-354.
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