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Cd14 beads

Manufactured by Miltenyi Biotec
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CD14 beads are magnetic microbeads coated with anti-human CD14 antibodies. They are used for the isolation and enrichment of CD14-positive cells, such as monocytes, from various biological samples.

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Lab products found in correlation

Ficoll paque, by GE Healthcare (2 mentions) Ficoll paque plus, by GE Healthcare (2 mentions) Cd56 beads, by Miltenyi Biotec (2 mentions) Lymphoprep, by Axis-Shield (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Gm csf, by Thermo Fisher Scientific (2 mentions) Sb203580, by Merck Group (1 mentions) Lps rs, by InvivoGen (1 mentions) Cu cpt22, by Bio-Techne (1 mentions) Rpmi 1640, by Corning (1 mentions) Fibronectin, by Corning (1 mentions) Fetal bovine serum (fbs), by Atlanta Biologicals (1 mentions) Gm csf, by Sanofi (1 mentions) Roswell park memorial institute (rpmi), by Corning (1 mentions) Interleukin 4 (il 4), by R&D Systems (1 mentions) Penicillin and streptomycin, by Biosera (1 mentions) Rpmi 1640 medium, by Biosera (1 mentions) Hela cell, by American Type Culture Collection (1 mentions) Dmem medium, by Biosera (1 mentions) Fetal calf serum (fcs), by Biosera (1 mentions)

Spelling variants of this product

Anti-CD14 magnetic beads (83 citations) Paramagnetic CD14-beads (6 citations) CD14 and CD16 beads (4 citations) CD14+ Magnetic Bead separation (3 citations) CD14+ bead selection (2 citations) CD14+ magnetic bead isolation (2 citations) CD14+ selection beads (2 citations) CD14-coupled magnetic beads (2 citations) CD14-mAb-coated micro beads (2 citations) CD14 antibody-coated magnetic beads (2 citations)

30 protocols using cd14 beads

1

Isolation of Human Microglia and GBM-Infiltrating Monocytes

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Human microglia were isolated as previously described53 . CD14+ GBM-infiltrating monocytes were extracted by mechanical disruption of ex vivo GBM samples collected from the operating room, and CD14+ cell isolation was performed by positive selection using CD14 beads (Miltenyi Biotec) after the homogenized tissue was passed through a cell strainer (70 μm). All patients had given informed consent for use of their resected brain tumor for research purposes. All studies using adult human tissue were approved by the McGill University Institutional Review Board (McGill University Health Centre Ethics Board; no. ANTJ2001/1). All experiments were conducted in accordance with the Helsinki Declaration. Peripheral blood mononuclear cells (PBMCs) from healthy controls (25–41 years old) were isolated using Ficoll gradient centrifugation. CD14+ cell purification was performed by positive selection using CD14 beads (Miltenyi Biotec) after homogenized tissue was passed through a cell strainer (70 μm). All procedures were approved by the Institutional Review Board of Brigham and Women’s Hospital.
Takenaka M.C., Gabriely G., Rothhammer V., Mascanfroni I.D., Wheeler M.A., Chao C.C., Gutiérrez-Vázquez C., Kenison J., Tjon E.C., Barroso A., Vandeventer T., de Lima K.A., Rothweiler S., Mayo L., Ghannam S., Zandee S., Healy L., Sherr D., Farez M.F., Prat A., Antel J., Reardon D.A., Zhang H., Robson S.C., Getz G., Weiner H.L, & Quintana F.J. (2019). Control of tumor-associated macrophages and T cells in glioblastoma via AHR and CD39. Nature neuroscience, 22(5), 729-740.
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2

Isolation of PBMCs and Immune Cell Subsets

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Peripheral blood mononuclear cells were purified from buffy coats originating from 450 ml blood of healthy blood donors (Uppsala Blood Transfusion Center, Uppsala University Hospital, Sweden), using Ficoll-Paque (GE Healthcare) density-gradient centrifugation. B cells, T cells and monocytes were isolated from dedicated batches of peripheral blood mononuclear cells, using positive selection with CD19+, CD3+ and CD14+ beads (Miltenyi Biotec), respectively, according to the manufacturer's instructions.
Allum F., Shao X., Guénard F., Simon M.M., Busche S., Caron M., Lambourne J., Lessard J., Tandre K., Hedman Å.K., Kwan T., Ge B., Rönnblom L., McCarthy M.I., Deloukas P., Richmond T., Burgess D., Spector T.D., Tchernof A., Marceau S., Lathrop M., Vohl M.C., Pastinen T, & Grundberg E. (2015). Characterization of functional methylomes by next-generation capture sequencing identifies novel disease-associated variants. Nature Communications, 6, 7211.
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3

Analyzing Inflammatory Signaling Pathways

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The reagents were obtained from the following sources: purified LPS from Escherichia coli strain 0111:B4 (Enzo Life Sciences), PamCSK4 from EMC microcollections, CD14 beads (MiltenyiBiotec), RPMI1640 (Cellgro), fetal bovine serum (FBS) (Atlanta Biologicals), penicillin-streptomycin (Invitrogen), and endotoxin free bovine serum albumin (BSA) (MP Biomedicals), Bronchial Epithelial Growth Medium BEGM bullet kit (Lonza), Bronchial Air Liquid Interface B-ALI media (Lonza), bovine collagen type I (Corning), fibronectin (Corning), LPS from Rhodobacter sphaeroides (RS-LPS) (Invivogen), CUCPT22 (Tocris Bioscience), SB203580 (Sigma-Aldrich) and JSH23 (Calbiochem). Scrambled siRNA control and siIκBζ (sequence UGAUGGACCUGCUUGCAAA) were purchased from Dharmacon Thermo Scientific. Rabbit antiserum against IκBζ was generated in our laboratory using recombinant protein expressed in E. coli [17 (link)]. Beta-actin antibody (monoclonal clone C4) and HSP 90α/β antibody (mouse monoclonal) were purchased from MP Biomedicals and Santa Cruz Biotechnology respectively.
Sundaram K., Rahman M.A., Mitra S., Knoell D.L., Woodiga S.A., King S.J, & Wewers M.D. (2016). IκBζ Regulates Human Monocyte Pro-Inflammatory Responses Induced by Streptococcus pneumoniae. PLoS ONE, 11(9), e0161931.
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4

Tracing Immune Cell Dynamics Post-Transplant

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Following hematopoietic recovery, and prior to (200–300 days posttransplant) or following SHIV challenge (300–400 days posttransplant, 100 days postinfection), large volume blood draws were collected by venipuncture into heparin collection tubes. Whole blood was separated by Ficoll centrifugation, and the mononuclear cell layer was subjected to bead-based positive selection with CD3, CD4, or CD14 beads (Miltenyi Biotec, Bergisch Gladbach, Germany). Subset purities were confirmed by flow cytometry, and the remainder of cells were processed for total genomic DNA for analysis of Cal-1 gene marking by Taqman.
Peterson C.W., Haworth K.G., Burke B.P., Polacino P., Norman K.K., Adair J.E., Hu S.L., Bartlett J.S., Symonds G.P, & Kiem H.P. (2016). Multilineage polyclonal engraftment of Cal-1 gene-modified cells and in vivo selection after SHIV infection in a nonhuman primate model of AIDS. Molecular Therapy. Methods & Clinical Development, 3, 16007-.
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5

Monocyte-derived Dendritic Cell Generation

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Monocyte-derived DCs were generated from peripheral blood mononuclear cells as described (29 (link)). Briefly, CD14+ monocytes were isolated from PBMCs by immunomagnetic bead selection using CD14 beads following the manufacturer’s protocol (Miltenyi Biotec). CD14+ cells were suspended in 1% healthy donor plasma in RPMI (Cellgro), supplemented with IL-4 (25μg/ml; R&D Systems) and GM-CSF (20ng/ml; Leukine Sargamostim, Genzyme) on days 0, 2 and 4 of culture. Immature monocyte-derived dendritic cells (Mo-DCs) were harvested on Days 5–6 and used for experiments described below. The CD14 negative fraction of PBMCs was cultured in the presence of 5% Pooled Human Serum (PHS, Labquip Ltd.) in RPMI. For some experiments BDCA3+ myeloid dendritic cells were isolated from the PBMCs using BDCA3 MACS beads (Miltenyi Biotec).
Sehgal K., Ragheb R., Fahmy T.M., Dhodapkar M.V, & Dhodapkar K.M. (2014). Nanoparticle-mediated combinatorial targeting of multiple human dendritic cell (DC) subsets leads to enhanced T cell activation via IL-15-dependent DC crosstalk. Journal of immunology (Baltimore, Md. : 1950), 193(5), 2297-2305.
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6

Culturing and Stimulating Immune Cells

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Human embryonic kidney (HEK) 293T cells (Unitech Ltd) and HeLa cells (ATCC) were cultured in DMEM medium (Biosera) supplemented with 10% FCS (Biosera), penicillin and streptomycin (100 µg/ml) (Biosera). Human PBMCs were isolated from whole blood by density gradient centrifugation using Ficoll-Paque Plus (GE Healthcare). Human monocytes were subsequently extracted from PBMCs by positive selection using CD14 beads (Miltenyi Biotec) and cultured in RPMI-1640 medium (Biosera) supplemented with 10% FCS and penicillin–streptomycin (100 µg/ml). Following stimulation, supernatants were collected and RANTES levels determined by ELISA (R&D Systems) according to the manufacturer's recommendations.
Wynne C., Lazzari E., Smith S., McCarthy E.M., Ní Gabhann J., Kallal L.E., Higgs R., Cryan S.A., Biron C.A, & Jefferies C.A. (2014). TRIM68 Negatively Regulates IFN-β Production by Degrading TRK Fused Gene, a Novel Driver of IFN-β Downstream of Anti-Viral Detection Systems. PLoS ONE, 9(7), e101503.
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7

Transwell Assay for Immune Cell Migration

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Blood samples from ICC patients were obtained from Liver Cancer Institute, Zhongshan Hospital. CD14+ cells were sorted by CD14+ beads (Miltenyi Biotec). Ten thousand sorted CD14+ cells were added to the upper chambers of Transwell (5 μm, 24-well format; Corning, USA). The supernatants from various fibroblasts were added to the lower chambers with or without CCL2 (100 ng/ml). Cells migrated to the lower chambers were counted by CyAn (Beckman Coulter) after 4 h incubation.
Lin Y., Li B., Yang X., Cai Q., Liu W., Tian M., Luo H., Yin W., Song Y., Shi Y, & He R. (2019). Fibroblastic FAP promotes intrahepatic cholangiocarcinoma growth via MDSCs recruitment. Neoplasia (New York, N.Y.), 21(12), 1133-1142.
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8

Monocyte Differentiation with M-CSF

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Human CD14+ monocytes were purified from human PBMCs of healthy donors using CD14+ beads (catalog number 130-050-201; Miltenyi Biotec), according to the manufacturer’s instructions. Purified CD14+ monocytes were cultured in minimal essential medium alpha (α-MEM) supplemented with 10% FBS and 20 ng/mL human recombinant macrophage colony-stimulating factor (M-CSF) (catalog number 574804; BioLegend). Fresh media with M-CSF were changed every 3 days until the cells reached 70 to 80% confluence (days 12 to 14).
Xiang J., Lu M., Shi M., Cheng X., Kwakwa K.A., Davis J.L., Su X., Bakewell S.J., Zhang Y., Fontana F., Xu Y., Veis D.J., DiPersio J.F., Ratner L., Sanderson R.D., Noseda A., Mollah S., Li J, & Weilbaecher K.N. (2022). Heparanase Blockade as a Novel Dual-Targeting Therapy for COVID-19. Journal of Virology, 96(7), e00057-22.
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9

Isolation of Primary Macrophages

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CD14 beads (Miltenyi Biotec, USA) were used to purify primary human and mouse macrophages, which were then cultured in GM_CSF containing medium as described under methods.
Khan A., Sayedahmed E.E., Singh V.K., Mishra A., Dorta-Estremera S., Nookala S., Canaday D.H., Chen M., Wang J., Sastry K.J., Mittal S.K, & Jagannath C. (2021). A recombinant bovine adenoviral mucosal vaccine expressing mycobacterial antigen-85B generates robust protection against tuberculosis in mice. Cell Reports Medicine, 2(8), 100372.
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10

Dengue Virus Modulation of B Cell Response

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Studies with human blood cells were pre-approved by the Emory University institutional review board. PBMC from total of 10 healthy donors were purified from blood by a standard technique using LSM medium (MPBiomedicals). Monocytes were isolated by the positive selection using CD14 beads (Miltenyi) and B cells were isolated using the CD19 beads (Miltenyi) to a purity of ≥95%. CD14+ monocytes or MDDC were infected with the DENV-2 (strain 16681) at MOI=1 for 48hr. Resting, allogenic CD19+ B cells were stained with CFSE (Invitrogen) and co-cultured in 96-well plates with the MDDC or monocytes at the ratio of 5:1 in the presence of 20U/ml of IL-2 (Peprotech) and 50nM CpG type B 2006 (TriLink Biotech) and blocking Ab. After 6 days sups were collected for IgG/A/M ELISA and cells were stained with the appropriate Ab cocktail and analyzed for CFSE dilution and B cell phenotype on a LSRII flow cytometer (BD). Detailed conditions of assays are described in the supplemental materials.
Kwissa M., Nakaya H.I., Onlamoon N., Wrammert J., Villinger F., Perng G.C., Yoksan S., Pattanapanyasat K., Chokephaibulkit K., Ahmed R, & Pulendran B. (2014). Dengue Virus Infection Induces Expansion of a CD14+CD16+ Monocyte Population that Stimulates Plasmablast Differentiation. Cell host & microbe, 16(1), 115-127.
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