Cd11b antibody

Manufactured by Miltenyi Biotec

Sourced in Germany

The CD11b antibody is a laboratory tool used to identify and study cells expressing the CD11b antigen, which is a protein found on the surface of certain immune cells. It can be used for flow cytometry and other immunological applications.

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Lab products found in correlation

Facsaria 3, by BD (2 mentions) Collagenase b 0.2, by Roche (2 mentions) Facscalibur, by BD (2 mentions) Gentlemacs octo dissociator with heater, by Miltenyi Biotec (1 mentions) Adult brain dissociation kit, by Miltenyi Biotec (1 mentions) Debris removal solution, by Miltenyi Biotec (1 mentions) Il 34, by Thermo Fisher Scientific (1 mentions) Collagenase 4, by Thermo Fisher Scientific (1 mentions) Glutamax, by Merck Group (1 mentions) Primaria 6 well plate, by Corning (1 mentions) M csf1, by Thermo Fisher Scientific (1 mentions) Albumax 1, by Thermo Fisher Scientific (1 mentions) Ascorbic acid, by Merck Group (1 mentions) Sodium chloride (nacl), by Merck Group (1 mentions) Gem21, by Gemini Bio (1 mentions) Ultra low attachment 6 well plate, by Corning (1 mentions) Macs smartstrainer 70 m, by Miltenyi Biotec (1 mentions) Macs c tube, by Miltenyi Biotec (1 mentions) Macs octo dissociator with heaters, by Miltenyi Biotec (1 mentions) Rneasy plus mini kit, by Qiagen (1 mentions)

Spelling variants of this product

CD11b (40 citations) PE-conjugated anti-CD11b antibodies (4 citations) Anti-CD11b antibody (4 citations) APC-CD11b antibody (2 citations) Anti-CD11b antibody- conjugated microbeads (2 citations) Magnetic CD11b antibody beads (2 citations) Anti-CD11b-PE conjugated primary antibody (2 citations) Anti-CD11b-FITC antibody (2 citations)

7 protocols using cd11b antibody

Isolation of Murine Microglia by MACS

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Brains were collected from mice for cell dissociation, and microglia were isolated by magnetic antibody-based cell sorting (MACS) using CD11B antibody (Miltenyi Biotec, Bergisch Gladbach, Germany, dilution 1:10) (details in Supplementary Methods) according to the manufacturer’s protocol using all recommended reagents and equipment. The purity of MACSed CD11B+ fractions was validated using FACS analysis of CD11B fluorescence, and the purity was further validated with RT-qPCR of the positive and negative CD11B cell fractions. Details in Supplementary Methods.

Krishnan M.L., Van Steenwinckel J., Schang A.L., Yan J., Arnadottir J., Le Charpentier T., Csaba Z., Dournaud P., Cipriani S., Auvynet C., Titomanlio L., Pansiot J., Ball G., Boardman J.P., Walley A.J., Saxena A., Mirza G., Fleiss B., Edwards A.D., Petretto E, & Gressens P. (2017). Integrative genomics of microglia implicates DLG4 (PSD95) in the white matter development of preterm infants. Nature Communications, 8, 428.

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Microglial Isolation and Purity Analysis

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Cortical samples were collected from CTRL and LPD/IL-1β pups at P1, P4, P7, P10, and P20. Primary microglial cells were sorted at P1, P4, P7, P10, and P20 using magnetic antibody-based cell sorting (MACS) and CD11b antibody (Miltenyi Biotec, Bergisch Gladbach, Germany), as previously reported [32 (link)]. The purity of the P4 and P7 MACS microglial cell sorting was validated by RT-qPCR performed on the positive and negative CD11b+ cell fractions for Itgam, Gfap, Neun, and Mbp and the arbitrary units normalized to the respective negative fraction in CTRL: P4 CTRL (mean ± SEM)—Itgam 99.78 ± 11.29, Gfap 0.13 ± 0.01, Neun 0.08 ± 0.02, and Mbp 0.02 ± 0.003; P4 LPD/IL-1β—Itgam 109.15 ± 5.27, Gfap 0.11 ± 0.1, Neun 0.02 ± 0.002, and Mbp 0.01 ± 0.01; P7 CTRL—Itgam 203.80 ± 9.95, Gfap 0.07 ± 0.01, Neun 0.04 ± 0.004, and Mbp 0.06 ± 0.02; and P7 LPD/IL-1β—Itgam 277.5 8 ± 23.27, Gfap 0.09 ± 0.01, Neun 0.05 ± 0.01, and Mbp 0.02 ± 0.003.

Zinni M., Mairesse J., Pansiot J., Fazio F., Iacovelli L., Antenucci N., Orlando R., Nicoletti F., Vaiman D, & Baud O. (2021). mGlu3 receptor regulates microglial cell reactivity in neonatal rats. Journal of Neuroinflammation, 18, 13.

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Isolation and Purification of Murine Microglia

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Briefly, spinal cord and brain from adult C57/Bl6 mice (8–10 weeks old) were removed and enzymatically digested with a collagenase B 0.2% (Roche Diagnostics GmbH) and trypsin-EDTA 0.2% at 37°C for 30 min, and then passed through a cell strainer of 40 μm (BD falcon), and cell suspension centrifuged twice at 300g for 10 minutes at 4°C. Microglia were isolated by magnetic sorting using a CD11b antibody (MiltenyiBiotec) and then stained with PerCP-Cy5.5-conjugated CD45 and PE-Cy7-conjugated CD11b antibodies for further purification on cell sorter (FACSARIATM III, BD Bioscience). Microglia cells were assessed on a flow cytometer (FACSCalibur; BD Biosciences), and only samples showing population >90% purity were used for gene expression analysis.

López-Serrano C., Santos-Nogueira E., Francos-Quijorna I., Coll-Miro M., Chun J, & López-Vales R. (2018). Lysophosphatidic acid receptor type 2 activation contributes to secondary damage after spinal cord injury in mice. Brain, behavior, and immunity, 76, 258-267.

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Brain Cell Dissociation and Enrichment

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Firstly, brains were dissociated into single-cell suspensions using the adult brain dissociation kit from Miltenyi (Bergisch Gladbach, Germany) (#130-107-677) based on a combination of enzymatic and mechanical dissociation protocol. Briefly, tissues are sliced into 8 sagittal sections and placed in tubes containing an enzyme cocktail. Each sample was then dissociated for 30 min at 37 °C on the gentleMACS Octo Dissociator with Heaters (#130-096-427, Miltenyi Biotec) using the 37C_ABDK_01 program. Then, myelin and debris were removed with Debris Removal solution (#130-109-398, Miltenyi Biotec). The resulting cell suspension was labeled with a CD11b antibody (#130-093-634) from Miltenyi or an O4 antibody (#130-096-670) and placed on a MACS^® column placed in the magnetic field of a MACS separator. CD11b⁺ cells were eluted, and their purity was validated by flow cytometry using the CD11b-FITC antibody (#130-113-796) or an O4-APC antibody (#130-115-810) from Miltenyi and the BD AccuriTM Cytometer. Cells were resuspended in Trizol and kept at −80 °C until further use.

Birmpili D., Charmarke Askar I., Pham-Van L.D., Kuntzel T., Spenlé C., Riou A, & Bagnard D. (2022). Toward a Combination of Biomarkers for Molecular Characterization of Multiple Sclerosis. International Journal of Molecular Sciences, 23(22), 14000.

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Microglia-like Cells from iPSCs

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We derived microglia-like cells from iPSCs as previously described (Muffat et al., 2016 (link)). Briefly, iPSCs on MEFs-coated plates were dissociated with Collagenase IV (Thermo Fisher Scientific). iPSCs were then resuspended in MGD media [Neurobasal media supplemented with 0.5X Gem21 (Gemini Bio-products), 0.5X Neuroplex N2 (Gemini Bio-products), 0.2% Albumax I (Thermo Fisher Scientific), 5 mM sodium chloride (Sigma-Aldrich), 1X sodium pyruvate, 1X P/S, 1X GlutaMAX, 3.5 ng/ml biotin (Sigma-Aldrich), 10 μM ascorbic acid (Sigma-Aldrich) and 1.7% lactic syrup (Sigma-Aldrich)] with 10 ng/ml IL-34 (Peprotech) and 10 ng/ml M-CSF1(Peprotech)], and cultured in ultra-low attachment 6-well plates (Corning). Once the phase-bright neutralized spheroids and cystic bodies appeared, Embryoid bodies were gently triturated to shear off cells of interest, and supernatants were transferred to a single well of Primaria 6-well plate (Corning). Attached cells showed morphological characteristics of microglia and microglia precursors. Cells from 6 consecutive productions were pooled and purified by FACS using CD11b antibody (Miltenyi Biotec). Collected cells were further maintained in MGD media with 100 ng/ml IL-34 and 5 ng/ml M-CSF for all experiments.

Lin Y.T., Seo J., Gao F., Feldman H.M., Wen H.L., Penney J., Cam H.P., Gjoneska E., Raja W.K., Cheng J., Rueda R., Kritskiy O., Abdurrob F., Peng Z., Milo B., Yu C.J., Elmsaouri S., Dey D., Ko T., Yankner B.A, & Tsai L.H. (2018). APOE4 causes widespread molecular and cellular alterations associated with Alzheimer’s disease phenotypes in human iPSC-derived brain cell types. Neuron, 98(6), 1141-1154.e7.

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Mouse Brain Dissociation and Cell Sorting

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Whole mouse brains were dissected from adult female CKp25 mice and washed in cold PBS with Ca²⁺ and Mg²⁺ (14287080; Thermo Fisher Scientific), placed on a Petri dish, finely diced with a razor, transferred to a MACS C Tube (130-093-237; Miltenyi), treated with the Adult Brain Dissociation Kit, mouse and rat (130-107-677; Miltenyi), and digested with a gentleMACS Octo Dissociator with Heaters (130-096-427; Miltenyi) for 30 min at 37°C. The slurry was then strained through a MACS 70 µm SmartStrainer (130-098-462; Miltenyi). Debris removal solution was then added, followed by a 3,000 ×g, 10 min, 4°C spin, discarding of two upper phases, and PBS wash step 1,000g, 10 min, 4°C spin. Cells were then labeled with CD11b antibody (130-110-554, 1:25 dilution; Miltenyi) and DAPI (D9542-1MG; Millipore) for 40 min followed by sorting into PBS with 1% BSA (700-100P; Gemini). Cells were then pelleted and subject to mRNA extraction using the RNeasy Plus Mini Kit (74134; Qiagen).

Ralvenius W.T., Mungenast A.E., Woolf H., Huston M.M., Gillingham T.Z., Godin S.K., Penney J., Cam H.P., Gao F., Fernandez C.G., Czako B., Lightfoot Y., Ray W.J., Beckmann A., Goate A.M., Marcora E., Romero-Molina C., Ayata P., Schaefer A., Gjoneska E, & Tsai L.H. (2023). A novel molecular class that recruits HDAC/MECP2 complexes to PU.1 motifs reduces neuroinflammation. The Journal of Experimental Medicine, 220(11), e20222105.

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Isolation of Murine Microglia

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Briefly, spinal cord and brain from adult C57/Bl6 mice (8–10 weeks old) were removed and enzymatically digested with a collagenase B 0.2% (Roche Diagnostics GmbH) and trypsine-EDTA 0.2% at 37 °C for 30 min, and then passed through a cell strainer of 40 μm mesh (BD falcon). Cell suspension was centrifuged twice at 300 g for 10 minutes at 4 °C, and microglial cells were first isolated by magnetic sorting using a CD11b antibody (MiltenyiBiotec) and then stained with PerCP-Cy5.5-conjugated CD45 and PE-Cy7-conjugated CD11b antibodies for further purification on cell sorter (FACSARIATM III, BD Bioscience). Microglia cells were assessed on a flow cytometer (FACSCalibur; BD Biosciences), and only populations presenting >90% purity were used for gene expression analysis.

Martínez-Muriana A., Mancuso R., Francos-Quijorna I., Olmos-Alonso A., Osta R., Perry V.H., Navarro X., Gomez-Nicola D, & López-Vales R. (2016). CSF1R blockade slows the progression of amyotrophic lateral sclerosis by reducing microgliosis and invasion of macrophages into peripheral nerves. Scientific Reports, 6, 25663.

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