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Mouse gm csf

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Mouse GM-CSF is a recombinant protein that functions as a cytokine, stimulating the production and differentiation of granulocytes and macrophages. It is commonly used in cell culture applications involving mouse-derived cells.

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Lab products found in correlation

Lsm 700 confocal microscope, by Zeiss (2 mentions) Red blood cell lysis buffer, by Merck Group (1 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by GE Healthcare (1 mentions) Macsquant analyzer, by Miltenyi Biotec (1 mentions) Female balb c mice, by Janvier Labs (1 mentions) Mouse il 4, by BioLegend (1 mentions) Cy3 streptavidin, by Jackson ImmunoResearch (1 mentions) Brefeldin a, by Thermo Fisher Scientific (1 mentions) Interleukin 2 (il 2), by Thermo Fisher Scientific (1 mentions) Anti β actin, by BioLegend (1 mentions) Anti cd3, by BioLegend (1 mentions) Pe anti mouse cd4, by BioLegend (1 mentions) Anti phospho stat3 tyr705, by BD (1 mentions) Alexa fluor 488 antimouse foxp3, by BioLegend (1 mentions) Fitc anti mouse cd11c, by BioLegend (1 mentions) Anti stat3, by Santa Cruz Biotechnology (1 mentions) Gm csf, by Thermo Fisher Scientific (1 mentions) Flt 3, by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by Lonza (1 mentions)

Close spelling variants of this product

LEGEND MAX Mouse GM-CSF ELISA Kit (2 citations) ELISA MAX Deluxe Set Mouse GM-CSF (2 citations) Mouse (Catalog 713704) and human (Catalog 713604) GM-CSF (2 citations) Recombinant mouse GM-CSF (27 citations) Recombinant Mouse GM-CSF (carrier-free) (2 citations) Mouse GM-CSF ELISA kit (3 citations) Recombinant mouse granulocyte-macrophage colony-stimulating factor (GM-CSF) (4 citations) Anti‐mouse GM‐CSF PE (2 citations) GM-CSF (148 citations)

6 protocols using mouse gm csf

1

Bone Marrow-Derived Dendritic Cell Generation

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Bone marrow cells from 6 weeks Balb/c female mice (Janvier) were extracted from leg bones with a 25G × 5/8 needle (Terumo). Red blood cells were eliminated by treatment with red blood cell lysis buffer (Sigma) at room temperature, following the manufacturer’s protocol. After centrifugation, the cells were washed three times with RPMI1640 (PAA Laboratories). The cells were then resuspended and grown for 10 days in RPMI 1640 medium containing 1 mM sodium pyruvate, 50 mM thioglycerol, 25 µg/mL penicillin streptomycin (Garamycin), 10% fetal calf serum (Gibco Life Technologies), and 50 ng/mL mouse GMCSF (Biolegend), in a humidified incubator at 37°C with 5% CO2.
Number and size of BMDCs cells were evaluated by incubation with alexa-fluor-488 anti-CD86 (Biolegend) and R-PE conjugated Monoclonal antibody specific to Mouse CD11c (invitrogen). Marking was analyzed by flow cytometry using a MACSQuant analyzer (Miltenyi Biotec), following the manufacturer’s instructions. Proportion above 90% of CD86 and CD11c positive cells were routinely obtained.
Christophe M., Kuczkowska K., Langella P., Eijsink V.G., Mathiesen G, & Chatel J.M. (2015). Surface display of an anti-DEC-205 single chain Fv fragment in Lactobacillus plantarum increases internalization and plasmid transfer to dendritic cells in vitro and in vivo. Microbial Cell Factories, 14, 95.
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2

Generating Bone Marrow-Derived DCs

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Bone marrow cells were flushed from the tibia of either IRAK-M−/− NOD or WT NOD mice. Cells were resuspended by repeated pipetting in culture medium, and erythrocytes were lysed before the culture. Bone marrow cells were then counted and seeded in 60-mm culture dishes at a density of 1.5 × 106/mL in DC differentiation medium (RPMI-1640 complete medium) with 5% FCS, 20 ng/mL mouse IL-4 (Biolegend), and 20 ng/mL mouse GM-CSF (Biolegend) for 7 days at 37°C. Culture medium was replenished every other day. Cells were harvested on day 7, were analyzed by flow cytometry after staining with CD11b and CD11c, and >90% of cells were CD11c+, which were considered to be bone marrow–derived DCs (BMDCs).
Tan Q., Majewska-Szczepanik M., Zhang X., Szczepanik M., Zhou Z., Wong F.S, & Wen L. (2014). IRAK-M Deficiency Promotes the Development of Type 1 Diabetes in NOD Mice. Diabetes, 63(8), 2761-2775.
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3

Visualization of CD11c+ Cells and PSA

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CD11c+ cells were enriched from the naïve mouse splenocytes by positive immunomagnetic selection. Adherent cells were treated with VLPV-PSA (5×105 IU/ml) or left untreated, and cultured in a chamber slide in a presence of 2×103 U/ml of mouse GM-CSF (BioLegend, San Diego, CA). Two hours post-infection, Brefeldin A (eBioscience, San Diego, CA) was added into chamber wells in the concentration 3 μg/ml. Cells were cultured for 24 h at 37°C, 5%CO2. Slides were fixed with acetone. CD11c and PSA expression was detected by immunofluorescent staining using biotinylated hamster anti-mouse CD11c (clone N418, eBioscience) and goat anti-human PSA (R&D Systems) abs for 1 h, RT followed by addition of Streptavidin-Cy3 and donkey anti-goat Cy2 abs (both Jackson Immunoresearch Laboratories, West Grove, PA) for 45 min, RT. Nuclei were counterstained by NucBlue Live Cell Stain reagent (Molecular Probes, Eugene, OR). Images were acquired on the Zeiss LSM700 confocal microscope (40x water objective) using ZEN software.
Riabov V., Tretyakova I., Alexander R.B., Pushko P, & Klyushnenkova E.N. (2015). Anti-Tumor Effect of the Alphavirus-based Virus-like Particle Vector Expressing Prostate-Specific Antigen in a HLA-DR Transgenic Mouse Model of Prostate Cancer. Vaccine, 33(41), 5386-5395.
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4

Molecular Mechanisms in Dendritic Cell Activation

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TCDD and DIM were purchased from Cambridge Isotope (Tewksbury, MA, USA) and Sigma-Aldrich (St. Louis, MO, USA), respectively. Human TGF-β and murine Flt3L and IL-2 were from eBioscience (San Diego, CA, USA). Mouse GM-CSF was from BioLegend (San Diego, CA, USA). Antibodies used in the present study included: anti-STAT3 and anti-phospho-STAT5 (Tyr 694), and anti-phospho-IκBα (Ser 32/36) from Santa Cruz Biotechnology (USA); anti-phospho-c-Src (Tyr416) and anti-STAT5 from Cell Signaling Technology (Danvers, MA, USA); anti-phospho-STAT3 (Tyr705) from BD Biosciences (Franklin Lakes, NJ, USA); FITC anti-mouse CD11c, Alexa Fluor 647 anti-mouse CD103, anti-CD3, anti-β actin, PE anti-mouse CD4, Alexa Fluor 488 anti-mouse FoxP3 from BioLegend.
Park J.H., Choi A.J., Kim S.J, & Jeong S.Y. (2015). 3,3'-Diindolylmethane Inhibits Flt3L/GM-CSF-induced-bone Marrow-derived CD103+ Dendritic Cell Differentiation Regulating Phosphorylation of STAT3 and STAT5. Immune Network, 15(6), 278-290.
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5

Generation of Bone Marrow-Derived Dendritic Cells

Cited 2 times
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BMDCs were generated as previously described (25 (link), 30 (link), 31 (link), 33 (link), 34 (link)). Briefly, femurs and tibiae were collected from freshly euthanized B6 mice, BM harvested, and single cell seeded at 2 × 105/ml. RPMI 1640 (Lonza) was supplemented with 10% FBS and 1% penicillin/streptomycin, plus 20 ng/ml mouse GM-CSF (all from BioLegend). For generation of CD103+ BMDCs, Flt-3 (100 ng/ml, eBioscience) was used instead of GM-CSF. On day 4, fresh GM-CSF (20 ng/ml) was added to both culture conditions.
Ahadome S.D., Mathew R., Reyes N.J., Mettu P.S., Cousins S.W., Calder V.L, & Saban D.R. (2016). Classical dendritic cells mediate fibrosis directly via the retinoic acid pathway in severe eye allergy. JCI Insight, 1(12), e87012.
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6

Metabolic Modulation of Immune Cells

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2-DG (Merck Millipore, Darmstadt, Germany), mouse GM-CSF (BioLegend, San Diego, CA, USA), phosphoenolpyruvate (PEP) monosodium salt hydrate (Sigma-Aldrich, Saint Louis, MO, USA), sodium IA (Sigma-Aldrich), sodium oxamate (Sigma-Aldrich), DPI (Cayman, Ann Arbor, MI, USA).
Jian S.L., Chen W.W., Su Y.C., Su Y.W., Chuang T.H., Hsu S.C, & Huang L.R. (2017). Glycolysis regulates the expansion of myeloid-derived suppressor cells in tumor-bearing hosts through prevention of ROS-mediated apoptosis. Cell Death & Disease, 8(5), e2779-.
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