Blood was layered onto Ficoll-Hypaque gradient (GE Healthcare, Waukesha, WI, USA) to collect cord blood mononuclear cells (CBMCs) or peripheral blood mononuclear cells (PBMCs). Monocytes were isolated by positive CD14 selection with magnetic microbeads (Miltenyi Biotec, Auburn, CA, USA). Preparations were routinely >95% pure as assessed by flow cytometry (23 (link)–26 (link)). Monocytes were cultured in tissue culture dishes at 0.4 × 106 cells/ml in RPMI media containing fresh 10% autologous platelet-poor plasma, supplemented with recombinant human (rh) IL-4 (50 ng/ml) and rh granulocyte-macrophage colony-stimulating factor (rhGM-CSF) (100 ng/ml) (R&D Systems, Minneapolis, MN, USA) with one replenishment of fresh media/cytokines at day 3. After 5–6 days, immature MoDCs were handled, as described previously (23 (link), 25 (link)).
Rh granulocyte macrophage colony stimulating factor rhgm csf
Manufactured by R&D Systems
Sourced in United States
Rh granulocyte-macrophage colony-stimulating factor (rhGM-CSF) is a recombinant human protein that stimulates the production and function of granulocytes and macrophages.
Automatically generated - may contain errors
Lab products found in correlation
Ficoll hypaque gradient, by GE Healthcare (1 mentions)
Magnetic microbead, by Miltenyi Biotec (1 mentions)
Escherichia coli bl21, by GE Healthcare (1 mentions)
Recombinant human rh il 4, by R&D Systems (1 mentions)
Rhtnf α, by R&D Systems (1 mentions)
Pgex 6p 1, by GE Healthcare (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Phorbol 12 myristate 13 acetate, by Merck Group (1 mentions)
Ionomycin, by Apollo Scientific (1 mentions)
2 protocols using rh granulocyte macrophage colony stimulating factor rhgm csf
1
Differentiation of Monocytes to Dendritic Cells
2
Recombinant rBmpA Protein Production
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rBmpA was produced in Escherichia coli BL21 (GE Healthcare, Chicago, IL, USA) using the bacterial expression vector pGEX-6P1 (GE Healthcare) and the following primers with EcoRI and XhoI restriction sites: Forward, 5′-ACGAATTCATGAATAAAATATTGTTGTTGA-3′ and reverse, 5′-AGCTCGTAAATAAATTCTTTAAGAAA-3′ (4 (link),5 (link)). Pure ISOF was provided by Dr Weimin Yang [School of Pharmaceutical Science and Yunnan Key Laboratory of Pharmacology for Natural Products, Kunming Medical University (KMU), Kunming, China] and reconstituted at 1×105 µM/ml in dimethyl sulfoxide (DMSO) for stock solution. LPS and phorbol-12-myristate-13-acetate (PMA) were purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany) and ionomycin was purchased from Apollo Scientific Ltd. (Stockport, UK). Recombinant human (rh)IL-4, rhTNF-α and rh-granulocyte macrophage colony-stimulating factor (rhGM-CSF) were purchased from R&D Systems, Inc. (Minneapolis, MN, USA).
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