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Murine gm csf

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Murine GM-CSF is a recombinant mouse granulocyte-macrophage colony-stimulating factor (GM-CSF) protein. GM-CSF is a cytokine that stimulates the production and differentiation of granulocytes and macrophages from bone marrow progenitor cells.

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Lab products found in correlation

Murine il 4, by R&D Systems (4 mentions) Fetal bovine serum (fbs), by Atlas Biologicals (3 mentions) Doxycycline, by Merck Group (3 mentions) Methylcellulose, by R&D Systems (3 mentions) Murine recombinant il3, by R&D Systems (3 mentions) Gm csf, by R&D Systems (2 mentions) Murine recombinant stem cell factor scf, by R&D Systems (2 mentions) Penicillin and streptomycin, by Thermo Fisher Scientific (2 mentions) Cell line nucleofector kit 5, by Lonza (2 mentions) Fetal bovine serum (fbs), by Gemini Bio (2 mentions) Bbl thioglycollate medium, by BD (2 mentions) Iscove modified dulbecco media (imdm), by Lonza (2 mentions) Methocult h4034, by STEMCELL (2 mentions) Rpmi 1640, by Lonza (2 mentions) Alpha minimum essential medium, by Corning (2 mentions) Lipopolysaccharides (lps), by Merck Group (2 mentions) Imdm medium, by Lonza (1 mentions) Rpmi 1640 medium, by Lonza (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Merck Group (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)

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GM-CSF (599 citations) Recombinant murine GM-CSF (28 citations)

23 protocols using murine gm csf

1

Colony Formation Assay for AML Cells

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tNM AML cells were cultured in RPMI-1640 medium (Lonza) with 10% fetal bovine serum (Atlas Biologicals) with or without 1 μg/mL doxycycline (Sigma) for 48 hours and then plated in IMDM medium (Lonza) with 30% fetal bovine serum (Atlas Biologicals), 1.275% methylcellulose (R&D Systems), and 2 ng/mL murine GM-CSF (R&D Systems). Human cell lines were treated with pathway inhibitors for 24 hours in standard culture medium and then harvested and plated in MethoCult H4034 Optimum CFC Medium (Stem Cell Technologies) according to manufacturer's recommendations. Colonies were scored after 7–14 days on an inverted microscope.
Eckfeldt C.E., Pomeroy E.J., Lee R.D., Hazen K.S., Lee L.A., Moriarity B.S, & Largaespada D.A. (2016). RALB provides critical survival signals downstream of Ras in acute myeloid leukemia. Oncotarget, 7(40), 65147-65156.
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2

Analyzing Influenza Virus Infection in B Cells

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B cells from normal mice were infected with LAIV or incubated with inactivated WSN (BPL treatment) at an MOI of 1 in RPMI 1640 medium containing 0.5 μg/ml TPCK-trypsin and incubated at 37 °C for 1 h. The infected cells were washed five times with PBS and labeled with CFSE (Merck) (1 μM CFSE for 10 min at room temperature, then washed twice with complete RPMI 1640 plus 10% FCS), then incubated with GM-CSF-cultured bone marrow-derived dendritic cells (BMDC) (ratio: 1:3) in DC culture medium (RPMI 1640 supplemented with 20 ng/mL murine GM-CSF (R&D Systems), 10% FBS, 50 μM 2-ME, 100 units/mL penicillin, and 100 μg/mL streptomycin) for 24 h at 37 °C. After incubation, DCs were isolated by M-pluriBead Cell Separation kit (pluriSelect) following the procedure from the company. The number of DCs with the green fluorescent signal was analyzed by flow cytometry, and viral proteins were analyzed by western blot41 (link). Some isolated DCs were incubated in DC culture medium for another 24 h at 37 °C for detection of virus replication.
Wu C.Y., Chuang H.Y, & Wong C.H. (2020). Influenza virus neuraminidase regulates host CD8+ T-cell response in mice. Communications Biology, 3, 748.
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3

Isolation of Murine Glial Cells

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Primary cultures of mixed murine glial cells were prepared as previously described using a modified protocol.38 ,48 (link) Briefly, brains from C57/BL6 P2 neonatal pups (purchased from Charles River, Saffron Walden, United Kingdom) were extracted, rolled across sterile filter paper to remove vasculature and meninges, and were mechanically dissociated. Cells were resuspended in 40 mL per 175 cm2 flask (Corning, Amsterdam, Netherlands) and were maintained in Dulbecco's Modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS) and 1% penicillin–streptomycin (Invitrogen, Sigma Aldrich, Gillingham, United Kingdom) and incubated at 37°C (5% CO2/95% O2). A week later, 5 ng/mL of murine GM-CSF (R&D systems, Abingdon, United Kingdom) in fresh media was added to the flasks to expand yields, and the cells were maintained for a further week.48 (link)Microglia were harvested by overnight shaking in an orbital shaker incubator (37°C, 180 rev/min), and the supernatant was collected from the flasks to obtain the dissociated microglia. Trypsin was subsequently added to lift the astrocytic layer. Microglia and astrocytes cells were plated as required for further experiments.
Nicol L.S., Thornton P., Hatcher J.P., Glover C.P., Webster C.I., Burrell M., Hammett K., Jones C.A., Sleeman M.A., Billinton A, & Chessell I. (2018). Central inhibition of granulocyte-macrophage colony-stimulating factor is analgesic in experimental neuropathic pain. Pain, 159(3), 550-559.
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4

Isolation and Transduction of Murine CML-LSC

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Animal studies were performed according to a protocol approved by the Animal Care and Use Committees of Northwestern University and Jesse Brown VA Medical Center. Bone marrow mononuclear cells were obtained from the femurs of Wt or Icsbp−/− C57/BL6 mice. Granulocyte/monocyte progenitor cells were cultured (2 × 105 cells per ml) in DME media supplemented with 10% fetal calf serum, 1% pen-strep, 10 ng/ml murine GM-CSF (R & D Systems Inc., Minneapolis, MN), 10 ng/ml murine recombinant IL-3 (R & D Systems Inc.) and 100 ng/ml of murine recombinant stem cell factor (Scf: R & D Systems Inc.) (referred to as myeloid progenitor cells in these studies). CD34+ cells were separated from the cultures using the Miltenyi magnetic bead system for extraction of RNA or proteins. These cells represent the CML-LSC in murine models during CP [43 (link)]. For some experiments, cells were transduced with retroviral vectors to express p210 Bcr-abl (in MiGR1 vector), DN-Gas2 (in MSCVpuro vector), or DN-Shp2 (in MSCVneo vector). Replication incompetent retroviral vectors were generated using the Ecopack packaging plasmid, as described [23 (link)].
Hjort E.E., Huang W., Hu L, & Eklund E.A. (2016). Bcr-abl regulates Stat5 through Shp2, the interferon consensus sequence binding protein (Icsbp/Irf8), growth arrest specific 2 (Gas2) and calpain. Oncotarget, 7(47), 77635-77650.
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5

Thioglycollate-elicited Macrophage Electroporation Protocol

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Thioglycollate-elicited macrophages were recovered 4 days after i.p. injection of 2 ml BBL thioglycollate medium, brewer modified (4%; BD Biosciences) by peritoneal lavage with 5 ml phosphate buffered saline (PBS). The peritoneal macrophages were cultured in DMEM cell culture medium (DMEM containing 10% FBS (Gemini Bio Products), 1% penicillin and streptomycin (Life Technologies) at 37°C and 95% air/5% CO2. For electroporation, 2×106 macrophages were electroporated with GFP mRNA only (2 μg), GFP mRNA (0.5 μg) + Nek7 mRNA (2 μg), or GFP mRNA (0.5 μg) + Nek7K64M mRNA (2 μg). Cell Line Nucleofector Kit V (Lonza) and program Y-001 were applied. Murine BMDMs were collected by flushing bone marrow cells from femurs and tibiae of mice. These cells were cultured for 7 days in DMEM cell culture medium containing 10% conditioned medium from L929 cells. For BMDCs, bone morrow cells were cultured in Petri dishes in 10 ml DMEM cell culture medium containing 10 ng/ml of murine GM-CSF (R&D Systems). On day 3 of culture, this was replaced with fresh GM-CSF medium. Loosely adherent cells were transferred to a fresh Petri dish and cultured for an additional 4 days.
Shi H., Wang Y., Li X., Zhan X., Tan M., Fina M., Su L., Pratt D., Bu C.H., Hildebrand S., Lyon S., Scott L., Quan J., Sun Q., Russell J., Arnett S., Jurek P., Chen D., Kravchenko V.V., Mathison J.C., Moresco E.M., Monson N.L., Ulevitch R.J, & Beutler B. (2015). NLRP3 activation and mitosis are mutually exclusive events coordinated by NEK7, a new inflammasome component. Nature immunology, 17(3), 250-258.
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6

Breast Cancer and Dendritic Cell Culture Protocol

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Breast cancer cells 4T1 (#CRL-2539) and MDA-MB-231 (#HTB-26) were purchased from American Type Culture Collection (ATCC) and cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and 1% penicillin + streptomycin (P/S) and passaged biweekly. JAWSII cells, an immortalized dendritic cell line derived from the bone marrow of p53−/− C57BL/6 mice (ATCC #CRl-11904), were grown in 10% FBS (ATCC, #30-2020) and Alpha Minimum Essential Medium (Corning, #10-022-CV), 1% P/S supplemented with 5 ng/ml murine GM-CSF (R&D Systems, 415-ML-050, Minneapolis, MN). All cell lines were cultured and maintained as previously described.37 (link), 38 (link)
Jenkins S.V., Nima Z.A., Vang K.B., Kannarpady G., Nedosekin D.A., Zharov V.P., Griffin R.J., Biris A.S, & Dings R.P. (2017). Triple-negative breast cancer targeting and killing by EpCAM-directed, plasmonically active nanodrug systems. NPJ Precision Oncology, 1, 27.
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7

Thioglycollate-elicited Macrophage Electroporation Protocol

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Thioglycollate-elicited macrophages were recovered 4 days after i.p. injection of 2 ml BBL thioglycollate medium, brewer modified (4%; BD Biosciences) by peritoneal lavage with 5 ml phosphate buffered saline (PBS). The peritoneal macrophages were cultured in DMEM cell culture medium (DMEM containing 10% FBS (Gemini Bio Products), 1% penicillin and streptomycin (Life Technologies) at 37°C and 95% air/5% CO2. For electroporation, 2×106 macrophages were electroporated with GFP mRNA only (2 μg), GFP mRNA (0.5 μg) + Nek7 mRNA (2 μg), or GFP mRNA (0.5 μg) + Nek7K64M mRNA (2 μg). Cell Line Nucleofector Kit V (Lonza) and program Y-001 were applied. Murine BMDMs were collected by flushing bone marrow cells from femurs and tibiae of mice. These cells were cultured for 7 days in DMEM cell culture medium containing 10% conditioned medium from L929 cells. For BMDCs, bone morrow cells were cultured in Petri dishes in 10 ml DMEM cell culture medium containing 10 ng/ml of murine GM-CSF (R&D Systems). On day 3 of culture, this was replaced with fresh GM-CSF medium. Loosely adherent cells were transferred to a fresh Petri dish and cultured for an additional 4 days.
Shi H., Wang Y., Li X., Zhan X., Tan M., Fina M., Su L., Pratt D., Bu C.H., Hildebrand S., Lyon S., Scott L., Quan J., Sun Q., Russell J., Arnett S., Jurek P., Chen D., Kravchenko V.V., Mathison J.C., Moresco E.M., Monson N.L., Ulevitch R.J, & Beutler B. (2015). NLRP3 activation and mitosis are mutually exclusive events coordinated by NEK7, a new inflammasome component. Nature immunology, 17(3), 250-258.
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8

Clonogenic Assay for AML Cells

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NRD and NRI AML cells were cultured in RPMI-1640 (Lonza, Basel, Switzerland) with 10% FBS (Atlas Biologicals, Fort Collins, CO, USA) with or without 1 μg/mL doxycycline (Sigma) for 48 hours or inhibitors for 24 hours and then plated in IMDM (Lonza) with 30% FBS, 1.275% methylcellulose (R&D Systems, Minneapolis, MN, USA), and 2 ng/mL murine GM-CSF (R&D Systems). Human AML cells were treated with inhibitors for 24 hours and then plated in MethoCult H4034 (Stem Cell Technologies, Vancouver, BC, Canada). Colonies were scored after 7-14 days on an inverted microscope.
Pomeroy E.J., Lee L.A., Lee R.D., Schirm D.K., Temiz N.A., Ma J., Gruber T.A., Diaz-Flores E., Moriarity B.S., Downing J.R., Shannon K.M., Largaespada D.A, & Eckfeldt C.E. (2016). Ras oncogene-independent activation of RALB signaling is a targetable mechanism of escape from NRAS(V12) oncogene addiction in acute myeloid leukemia. Oncogene, 36(23), 3263-3273.
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9

Knockdown of Immune Regulators in Murine Bone Marrow Cells

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Single-cell suspensions of bone marrow cells were cultured in RPMI 1640 medium containing 10% heat-inactivated fetal calf serum and 1% penicillin-streptomycin supplemented with murine GM-CSF at a final concentration of 25 ng/ml (R&D Systems, Minneapolis, MN). Fresh GM-CSF was provided on day 4 of culture. The day-6 cells were harvested to knock down DHX15, MAVS and STING by shRNA. The knockdown efficiency was detected with western blotting 36 hours post shRNA treatment.
Lu H., Lu N., Weng L., Yuan B., Liu Y.J, & Zhang Z. (2014). DEAH (Asp-Glu-Ala-His) box helicase 15 senses double-stranded RNA in myeloid dendritic cells. Journal of immunology (Baltimore, Md. : 1950), 193(3), 1364-1372.
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10

Bone Marrow-Derived Dendritic Cell Culture

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Bone marrow-derived DCs were generated using a modified version of the protocol originally described by Inaba et al. [24 (link)], without lymphocyte depletion. Briefly, equal numbers of BM cells from WT and KIKO mice were suspended in complete RPMI supplemented with 20ng/mL murine GM-CSF and 20ng/mL murine IL-4 (R&D systems, Minneapolis, MN) and cultured in T75 flasks at 1 × 106 cells/ml (~20 × 106/flask) for 7 days. BMDCs were harvested from flasks, counted, and re-plated in 6-well plates at 1 × 106 cells/ml (4 × 106/well) for 18h. For spleen cells, mice were sacrificed and spleens harvested and kept in ice-cold RPMI. Spleens were processed and subjected to red blood cell lysis. Cells were washed twice in cold RPMI before being counted and cultured in 12- or 6-well plates at 1 × 106 cells/ml (2-4 × 106/well) for 18h.
Cunningham M.A., Wirth J.R., Naga O., Eudaly J, & Gilkeson G.S. (2014). Estrogen Receptor Alpha Binding to ERE is Required for Full Tlr7- and Tlr9-Induced Inflammation. SOJ immunology, 2(1), 07.
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