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Recombinant gm csf

Manufactured by BioLegend
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Recombinant GM-CSF is a cytokine that stimulates the production and function of granulocytes and macrophages. It is produced using recombinant DNA technology.

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Lab products found in correlation

Alexa fluor 488 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Penicillin, by Lonza (1 mentions) Streptomycin, by Lonza (1 mentions) L glutamine, by Lonza (1 mentions) Ionomycin, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Lonza (1 mentions) 2 3 cgamp, by InvivoGen (1 mentions) Dbs100, by American Type Culture Collection (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Penicillin, by Merck Group (1 mentions) Streptomycin, by Merck Group (1 mentions) Xav939, by Merck Group (1 mentions) Anti siglec f microbeads, by Miltenyi Biotec (1 mentions) Flt3l, by BioLegend (1 mentions) Macs ls column, by Miltenyi Biotec (1 mentions)

Close spelling variants of this product

Recombinant mouse granulocyte-macrophage colony-stimulating factor (GM-CSF) (4 citations) Recombinant murine GM-CSF (3 citations) Recombinant human GM-CSF (8 citations) Recombinant Mouse GM-CSF (carrier-free) (2 citations) Recombinant mouse GM-CSF (27 citations) GM-CSF (148 citations)

4 protocols using recombinant gm csf

1

Phenotyping Immune Cell Subsets by Flow Cytometry

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All of the materials used were purchased from Sigma-Aldrich (Saint Louis, MO, USA), unless otherwise indicated. Recombinant GM-CSF was purchased from BioLegend (San Diego, CA, USA) and Ionomycin from EMD Millipore (Burlington, MA, USA). TSP-1 derived peptide KRFK and inactive control peptide Lys-Gln-Phe-Lys (KQFK) were synthesized by Bionova (Madrid, Spain). Complete RPMI-1640 was prepared by adding 10 mM HEPES, 0.1 mM NEAA, 1 mM sodium pyruvate, 100 U/mL Penicillin, 100 mg/mL streptomycin, 200 mM l-glutamine (all from Lonza, Basel, Switzerland), and 10% fetal bovine serum (Atlanta Biologicals, GA, USA). Anti-CD4-PE-Cy-5 (clone RM4-5) was purchased from BioLegend (San Diego, CA, USA), anti-IFN-γ-FITC (clone XMG1.2), anti-IL-17-PE (clone eBio17B7), and anti-Foxp3-PE-Cy5 (clone FJK-16s) were purchased from eBioscience/Thermofisher scientific (San Diego, CA, USA). Anti-SMA (clone 1A4) was purchased from Abcam (Cambridge, MA, USA). Alexa Fluor 488-conjugated secondary antibodies were purchased from Thermofisher scientific (Waltham, MA, USA).
Soriano-Romaní L., Contreras-Ruiz L., López-García A., Diebold Y, & Masli S. (2018). Topical Application of TGF-β-Activating Peptide, KRFK, Prevents Inflammatory Manifestations in the TSP-1-Deficient Mouse Model of Chronic Ocular Inflammation. International Journal of Molecular Sciences, 20(1), 9.
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2

Generating Murine Dendritic Cells

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Bone marrow (BM) cells were isolated by flushing the long bones with PBS. BM-derived GM-CSF DC cultures were grown in 24 well plates as previously described57 (link) in RPMI 1640 supplemented with 40 pg/ml recombinant GM-CSF (BioLegend) and 5% FBS, plus 100 μg/ml penicillin, 100 μg/ml streptomycin, 2 mM L-glutamine, 10 mM HEPES, 1 nM sodium pyruvate, 1× MEM nonessential amino acids, and 2.5 μM β-mercaptoethanol (all Sigma). After 5 days of culture, BMDCs were harvested and replated with MOI (multiplicity of infection) =10 of Citrobacter rodentium (DBS100, ATCC) or incubated with 5 μg/ml of the 2’3’-cGAMP (cyclic [G(2’,5’)pA(3’,5’)p]) (Invivogen). For infection, bacteria were grown to exponential phase in lysogeny broth (LB) medium, then washed and resuspended in PBS.
Two hours post-treatment, culture medium was removed and cells resuspended in 1 ml of TRIzol Reagent (Life Technologies).
Mota-Meira M., Lapointe G., Lacroix C, & Lavoie M.C. (2000). MICs of Mutacin B-Ny266, Nisin A, Vancomycin, and Oxacillin against Bacterial Pathogens. Antimicrobial Agents and Chemotherapy, 44(1), 24-29.
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3

Isolation and Culture of Alveolar Macrophages

Cited 1 time
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Lung tissue was digested as described above to obtain a single-cell suspension. SiglecF+ cells were isolated using anti-SiglecF microbeads (Miltenyi Biotec) and plated in non-treated 6-well plates in RPMI 1640 media containing 1x GlutaMAX, 1x pyruvate, 1x penicillin/streptomycin, 10% fetal bovine serum (FBS), and 20 ng/ml recombinant GM-CSF (Biolegend), as previously described (31 (link)). Culture medium was replaced after 16 h of incubation at 37°C and again every 2 days in culture. Cells were collected on days 1 or 6 after plating. Where indicated, AMs were treated overnight with 5 μM of the Wnt/β-catenin inhibitor XAV939 (Sigma) or 1 μM of the Stat3 inhibitor Napabucasin (Fisher Scientific) before cells were harvested.
Scortegagna M., Du Y., Bradley L.M., Wang K., Molinolo A., Ruppin E., Murad R, & Ronai Z.A. (2023). Ubiquitin ligases Siah1a/2 control alveolar macrophage functions to limit carcinogen-induced lung adenocarcinoma. Cancer research, 83(12), 2016-2033.
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4

Dendritic Cell Differentiation from Murine Bone Marrow

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Femurs and tibias of mice were removed by dissection and attached muscle were removed. Bones were soaked in 70% ethanol for 5 min. The epiphyses were cut using a scalpel and the bone marrow was flushed out using a syringe and needle with basal culture medium (RPMI 1640 supplemented with 10% v/v fetal bovine serum (FBS), 50 μM 2-mercaptoethanol, 2 mM L-glutamine, and 1% v/v Penicillin/Streptomycin). Cells were centrifuged at 300×g for 5 min and re-suspended in ACK lysis buffer for 3 min at room temperature. Cells were passed through a 70 μm filter, centrifuged and re-suspended in basal culture medium and factors were added as indicated in results. The factors used to optimize differentiation of bone marrow cells into dendritic cells (DCs) with an intestinal phenotype were 200 ng/ml recombinant FMS-like tyrosine kinase 3 ligand (Flt3L, Biolegend), 20 ng/ml recombinant GM-CSF (Biolegend), and 1 μM all-trans-Retinoic acid (RA, Sigma). Cells were cultured at 37°C, 5% v/v atmospheric CO2 for 8 days before purification for CD11c+ cells by positive selection using MACS LS columns (Miltenyi) according to the manufacturer’s protocol.
Johnston L.J., Barningham L., Campbell E.L., Cerovic V., Duckworth C.A., Luu L., Wastling J., Derricott H, & Coombes J.L. (2023). A novel in vitro model of the small intestinal epithelium in co-culture with ‘gut-like’ dendritic cells. Discovery Immunology, 2(1), kyad018.
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