Anti cd11c beads

Manufactured by Miltenyi Biotec

Sourced in France

Anti-CD11c beads are magnetic beads that are coated with antibodies specific to the CD11c antigen. CD11c is a cell surface marker expressed on dendritic cells and other antigen-presenting cells. These beads can be used to isolate and enrich CD11c-positive cells from various samples, such as blood, tissues, or cell cultures. The beads facilitate the separation and purification of these cell populations for further analysis or experimental applications.

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Lab products found in correlation

Anti cd43 beads, by Miltenyi Biotec (2 mentions) Anti biotin bead, by Miltenyi Biotec (1 mentions) Ack buffer, by Thermo Fisher Scientific (1 mentions) Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (1 mentions) Rneasy plus micro kit, by Qiagen (1 mentions) Macs bead, by Miltenyi Biotec (1 mentions) Facsaria 2 sorter, by BD (1 mentions) Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions) Gm csf, by Thermo Fisher Scientific (1 mentions) Anti ifnγ xmg1.2, by Thermo Fisher Scientific (1 mentions) Hoechst, by Thermo Fisher Scientific (1 mentions) Anti cd43, by Miltenyi Biotec (1 mentions) Cd4 t cell isolation kit, by Miltenyi Biotec (1 mentions) Collagenase type 4, by Worthington (1 mentions) Ack buffer, by Lonza (1 mentions) Anti pe magnetic beads, by Miltenyi Biotec (1 mentions) Anti pe beads, by Miltenyi Biotec (1 mentions) Debris removal solution, by Miltenyi Biotec (1 mentions)

Close spelling variants of this product

Paramagnetic bead-conjugated anti-CD11c Anti-CD11c immunomagnetic bead purification Bead-conjugated anti-CD11c Bead-conjugated anti-CD11c mAb Anti-CD11c magnetic beads Anti-CD11c-coated magnetic beads Anti-CD11c MACS beads Anti-CD11c-conjugated magnetic beads Anti-mouse CD11c magnetic beads Anti-CD11c immunomagnetic beads

7 protocols using anti cd11c beads

Isolation of Immune Cell Subsets from Murine Spleen

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To isolate DCs, spleens were collected, incubated for 30 min at 37 °C in HBSS (Gibco) supplemented with CaCl₂, MgCl₂, and collagenase D at 400 U ml⁻¹ (Roche). After digestion, tissue was forced 5 times through a 21-gauge needle and filtered through a 70 μm strainer into a 15 ml falcon tube with PBS supplemented with 0.5% BSA and 2 mM EDTA (PBE). Red-blood cells were lysed with ACK buffer (Gibco), and the resulting cell suspensions were filtered through a 70-μm mesh into PBE. DCs were obtained by magnetic cell separation (MACS) using anti-CD11c beads (Miltenyi Biotec), as per the manufacturer’s instructions. To isolate CD4⁺ T cells, CD8⁺ T cells, and B cells, spleens were processed as above, without collagenase digestion. CD4⁺ T cells and CD8⁺ T cells were isolated by negative selection using a cocktail of biotinylated antibodies targeting Ter119, CD11c, CD11b, CD25, B220, NK1.1, and either CD8 (for CD4⁺ isolation) or CD4 (for CD8⁺ isolation), followed by anti-biotin beads (Miltenyi Biotec), as per the manufacturer’s instructions. B cells were obtained by negative selection using anti-CD43 beads (Miltenyi Biotec), as per the manufacturer’s instructions.

Nakandakari-Higa S., Canesso M.C., Walker S., Chudnovskiy A., Jacobsen J.T., Bilanovic J., Parigi S.M., Fiedorczuk K., Fuchs E., Bilate A.M., Pasqual G., Mucida D., Pritykin Y, & Victora G.D. (2023). Universal recording of cell–cell contacts in vivo for interaction-based transcriptomics. bioRxiv.

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Characterization of Splenic CD11c+ Cells

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CD11c⁺CD3e⁻B220⁻CD317⁻ splenic cells from immunized mice were first magnetically sorted with anti-CD11c-Beads (Miltenyi Biotec, Paris, France) and then by FACS ARIA after labeling with anti-CD11c/eF450 (N418), anti-B220/PE (RA3–6B2), anti-CD317/APC (eBio927) and anti-CD3e/FITC (17A2) antibodies. The purity was always > 96.3% of live cells. RNA from purified CD11c⁺ cells was extracted from cell lysates with the RNeasy Plus microkit (QIAGEN, Courtaboeuf, France). cDNA was generated using a RT2 First Strand Kit and quantified using the RT2 Profiler Mouse antiviral responses and NF-κB signaling pathway PCR arrays (SABiosciences, QIAGEN, Courtaboeuf, France), according to the manufacturer’s instructions. The results were analyzed using the SA Biosciences Data Analysis Web Portal (http://pcrdataanalysis.sabiosciences.com/pcr/arrayanalysis.php) and expressed as fold regulation compared to PBS-treated mice.

Cousin C., Oberkampf M., Felix T., Rosenbaum P., Weil R., Fabrega S., Morante V., Negri D., Cara A., Dadaglio G, & Leclerc C. (2019). Persistence of Integrase-Deficient Lentiviral Vectors correlates with the induction of STING-independent CD8+ T cell responses. Cell reports, 26(5), 1242-1257.e7.

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Purification of Lymphocyte Subsets

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Non-CD4⁺ lymph node and spleen T cells were removed by MACS beads (Miltenyi Biotec) after coating with biotin anti-CD8α, DX5, B220, CD3, CD11b, Ly-6G, and Ter119. Cells were further purified with a FACSAria 2 sorter (BD) to >97%. Spleen CD11c⁺ DCs were partially enriched with anti-CD11c beads (Miltenyi Biotec) and, where indicated, purified with a FACSAria 2 (BD) cell sorter as CD11c^highCD19⁻CD3⁻DX5⁻ DCs (>95%).

Sela U., Park C.G., Park A., Olds P., Wang S., Steinman R.M, & Fischetti V.A. (2016). Dendritic Cells Induce a Subpopulation of IL-12Rβ2-Expressing Treg that Specifically Consumes IL-12 to Control Th1 Responses. PLoS ONE, 11(1), e0146412.

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Bone Marrow-Derived DC-B Cell Co-Culture

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Bone marrow (BM)-derived DCs (BMDCs) were differentiated with 10 ng/ml GM-CSF (Peprotech) and 5 ng/ml IL-4 (Peprotech), then purified with anti-CD11c beads (Miltenyi Biotec), as previously described [3] (link). CD43⁻ B cells were isolated from B6 splenocytes by negative selection (Miltenyi Biotec). For BMDC-B cell co-cultures, 2×10⁴ BMDCs from either TC or B6 mice were cultured for 5 d with 10⁵ B cells and 10 ug/ml anti-CD40 Ab (1C10) (eBioscience) in duplicate as previously described [3] (link). The effect of cytokine inhibition on B cell proliferation was studied by adding 10 ug/ml of anti-IL-6 (MP5-20F3) (BD Biosciences) and/or anti-IFN-γ (XMG1.2) (eBioscience) to the co-cultures. To quantify cytokine secretion by activated BMDCs, 2×10⁴ BMDCs were cultured with 10 ug/ml anti-CD40 Ab for 24 h, after which RNA was extracted for qRT-PCR and gene array analyses, and supernatants were collected from the cultures for ELISA. IFN-γ was also quantified by intracellular flow cytometry, and BMDCs stimulated with either IL-12 (5 ng/ml) or IL-18 (20 ng/ml) were used as positive controls. Mice used in these experiments were 2–3 months of age.

Sang A., Zheng Y.Y., Yin Y., Dozmorov I., Li H., Hsu H.C., Mountz J.D, & Morel L. (2014). Dysregulated Cytokine Production by Dendritic Cells Modulates B Cell Responses in the NZM2410 Mouse Model of Lupus. PLoS ONE, 9(8), e102151.

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Analyzing Leishmania Infection in Dendritic Cells

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CD11c⁺ dendritic cells (DCs) were isolated using anti-CD11c beads (Miltenyi Biotec) [19 (link)]. BMDC were generated as previously described [91 (link)] and seeded at 10⁶ cells/ml onto 24-well plates. Cells were subsequently infected with PKH26-stained L. donovani amastigotes (Sigma, staining done according to manufacturer’s instructions) at a MOI of 5:1. Infected cells were then harvested 24, 48, and 72 h later, fixed with 2% PFA, stained with Hoechst (Invitrogen, staining done according to manufacturer’s instructions) and cytospined on PBS/ 1% BSA-prepared slides. Mounted slides were than analyzed using a fluorescent microscope, to evaluate parasite survival and target infection rate.

Hammami A., Charpentier T., Smans M, & Stäger S. (2015). IRF-5-Mediated Inflammation Limits CD8+ T Cell Expansion by Inducing HIF-1α and Impairing Dendritic Cell Functions during Leishmania Infection. PLoS Pathogens, 11(6), e1004938.

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Isolation of Immune Cell Populations

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To isolate DCs, spleens were harvested, incubated 30 min at 37 °C in RPMI, 2% FBS, 20 mM HEPES, 400 U/ml type IV collagenase (Worthington Biochemical Corporation) and disrupted to generate single cell suspensions. Red blood cells were lysed with ACK buffer (Lonza), and the resulting cell suspensions were filtered through a 70 μm mesh into PBS supplemented with 0.5% BSA and 2 mM EDTA (PBE). DCs were obtained by magnetic cell separation (MACS) using anti-CD11c beads (Miltenyi Biotec), as per manufacturer’s instructions. To isolate CD4⁺ T cells and B cells, spleens were processed as above, except for collagenase digestion, which was not performed. CD4⁺ T cells were isolated using CD4⁺ T cell isolation kit (Miltenyi Biotec) while B cells were obtained by negative selection using anti-CD43 beads (Miltenyi Biotec), as per manufacturer’s instructions. To isolate Igλ⁺ B cells form B1-8^hi mice, B cells were stained with anti-Igκ PE antibody and subsequently purified by negative selection using a combination of anti-CD43 and anti-PE magnetic beads (Miltenyi Biotec).

Pasqual G., Chudnovskiy A., Tas J.M., Agudelo M., Schweitzer L.D., Cui A., Hacohen N, & Victora G.D. (2018). Monitoring T cell–dendritic cell interactions in vivo by intercellular enzymatic labeling. Nature, 553(7689), 496-500.

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Hepatic Non-Parenchymal Cell Isolation

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Hepatic NPCs were isolated as described above and samples were then cleared with Debris Removal Solution according to the manufacturer's protocol (Miltenyi), followed by blocking with anti-CD16/32 antibody. For negative selection of DCs, NPCs were incubated with lineage-specific antibodies (F4/80-PE; CD3-PE; CD19 PE; NK1.1-PE) and Anti-PE-Beads (Miltenyi) to facilitate the retention of these cell lineages on LD columns according to the manufacturer's protocol. The cells in the flowthrough were incubated with Anti-CD11c-Beads (Miltenyi), and positive selection of CD11c+ cells with MS columns was performed according to the manufacturer's protocol (Miltenyi) .

Müller A.L., Casar C., Preti M., Krzikalla D., Gottwick C., Averhoff P., Rosenstiel P., Gelderblom M., Altfeld M., Lohse A.W., Steinmann S., Sebode M., Krause J., Schwinge D., Schramm C., Carambia A, & Herkel J. (2022). Inflammatory type 2 conventional dendritic cells contribute to murine and human cholangitis. Journal of hepatology, 77(6).

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