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Human macrophage colony stimulating factor

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Human macrophage colony-stimulating factor is a recombinant protein that functions as a cytokine. It is involved in the regulation and differentiation of macrophages, which are important immune cells.

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Lab products found in correlation

Monocyte isolation kit 2, by Miltenyi Biotec (2 mentions) Human granulocyte macrophage colony stimulating factor hgm csf, by Merck Group (2 mentions) Hm csf, by Merck Group (2 mentions) Hgm csf, by Merck Group (2 mentions) Facs canton 2, by BD (2 mentions) Lipopolysaccharides (lps), by Merck Group (2 mentions) Monocyte attachment medium, by PromoCell (2 mentions) Sepmate 50 system, by STEMCELL (1 mentions) 96 well plate, by Thermo Fisher Scientific (1 mentions) Rat bone marrow macrophages, by ScienCell (1 mentions) X vivo 15, by Lonza (1 mentions)

4 protocols using human macrophage colony stimulating factor

1

Isolation, Differentiation, and Culture of Human and Rat Macrophages

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Human monocytes were isolated from whole blood using the SepMate-50 system (Stemcell Technologies, Seattle, WA, USA). The protocol was approved by the University of Arizona IRB. Cells were then cryopreserved. Peripheral blood mononuclear cells (PBMCs) from normal (healthy) adult controls were placed in complete RPMI (RPMI, 10% fetal bovine serum, penicillin/streptomycin, and DNase (3 ng/mL)) and thawed to 37 °C. Cells were collected by centrifugation, resuspended in X-VIVO-15 (Lonza, Walkersville, MD, USA), a serum-free medium, plated into 96-well plates (Fisher Scientific) (105 cells in 100 μL medium/well), and placed in an incubator overnight at 37 °C with 5% CO2. On the following day PBMCs were washed with serum-free medium and resuspended in fresh medium, human Macrophage Colony Stimulating Factor (Sigma-Aldrich, St. Louis, MO, USA) was added to each well (1 ng/mL), and cells incubated for 5 days at 37 °C with 5% CO2 [5 (link)]. Rat bone marrow macrophages (ScienCell Research Laboratories, Carlsbad, CA, USA) were treated in the same manner with the exception that 10% rat serum was provided in the RPMI as per manufacturer’s directions.
Klotz S.A., Bradley N, & Lipke P.N. (2022). Blocking Serum Amyloid-P Component from Binding to Macrophages and Augmenting Fungal Functional Amyloid Increases Macrophage Phagocytosis of Candida albicans. Pathogens, 11(9), 1000.
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2

Monocyte Isolation and Macrophage Differentiation

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Monocytes were purified from PBMCs by negative selection (Monocyte Isolation Kit II; Miltenyi Biotec). Monocytes were suspended in monocyte attachment medium (PromoCell) and seeded at a density of 150,000/cm2 for 2 hours. Monocytes were co-stained with APC-Cy7-conjugated anti-CD11b, PE-Cy7-conjugated CD11c, Fitc conjugated anti-CD14, APC-conjugated CD16 or relative isotype control and then were analyzed using a FACS Canton II (BD Biosciences).
Primary human monocytes were differentiated into M1 macrophages using 20 ng/mL human Granulocyte-macrophage colony-stimulating factor (hGM-CSF, Sigma) or into M2 macrophages using 100 ng/mL human macrophage colony-stimulating factor (hM-CSF, Sigma) for 10 days. hGM-CSF differentiated M1 macrophages were stimulated with or without LPS (Sigma) 100 ng/mL for 6 hours.
Zhou Q., Wang H., Schwartz D.M., Stoffels M., Park Y.H., Zhang Y., Yang D., Demirkaya E., Takeuchi M., Tsai W.L., Lyons J.J., Yu X., Ouyang C., Chen C., Chin D.T., Zaal K., Chandrasekharappa S.C., Hanson E.P., Yu Z., Mullikin J.C., Hasni S.A., Wertz I., Ombrello A.K., Stone D.L., Hoffmann P., Jones A., Barham B.K., Leavis H.L., van Royen-Kerkof A., Sibley C., Batu E.D., Gül A., Siegel R.M., Boehm M., Milner J.D., Ozen S., Gadina M., Chae J., Laxer R.M., Kastner D.L, & Aksentijevich I. (2015). Loss-of-function mutations in TNFAIP3 leading to A20 haploinsufficiency cause an early onset autoinflammatory syndrome. Nature genetics, 48(1), 67-73.
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3

Monocyte Isolation and Macrophage Differentiation

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Monocytes were purified from PBMCs by negative selection (Monocyte Isolation Kit II; Miltenyi Biotec). Monocytes were suspended in monocyte attachment medium (PromoCell) and seeded at a density of 150,000/cm2 for 2 hours. Monocytes were co-stained with APC-Cy7-conjugated anti-CD11b, PE-Cy7-conjugated CD11c, Fitc conjugated anti-CD14, APC-conjugated CD16 or relative isotype control and then were analyzed using a FACS Canton II (BD Biosciences).
Primary human monocytes were differentiated into M1 macrophages using 20 ng/mL human Granulocyte-macrophage colony-stimulating factor (hGM-CSF, Sigma) or into M2 macrophages using 100 ng/mL human macrophage colony-stimulating factor (hM-CSF, Sigma) for 10 days. hGM-CSF differentiated M1 macrophages were stimulated with or without LPS (Sigma) 100 ng/mL for 6 hours.
Zhou Q., Wang H., Schwartz D.M., Stoffels M., Park Y.H., Zhang Y., Yang D., Demirkaya E., Takeuchi M., Tsai W.L., Lyons J.J., Yu X., Ouyang C., Chen C., Chin D.T., Zaal K., Chandrasekharappa S.C., Hanson E.P., Yu Z., Mullikin J.C., Hasni S.A., Wertz I., Ombrello A.K., Stone D.L., Hoffmann P., Jones A., Barham B.K., Leavis H.L., van Royen-Kerkof A., Sibley C., Batu E.D., Gül A., Siegel R.M., Boehm M., Milner J.D., Ozen S., Gadina M., Chae J., Laxer R.M., Kastner D.L, & Aksentijevich I. (2015). Loss-of-function mutations in TNFAIP3 leading to A20 haploinsufficiency cause an early onset autoinflammatory syndrome. Nature genetics, 48(1), 67-73.
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4

NLRP3-Mediated Macrophage Activation

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Bone marrow-derived macrophages (BMDM) were prepared by isolating bone marrow from the femurs of 8- to 10-week-old C57BL/6 or Nlrp3 KO mice, followed by incubation for 7–10 days with L-cell conditioned medium for macrophage differentiation as we described previously [20 (link)]. For NLRP3 inflammasome stimulation, differentiated BMDM were primed with LPS (100 ng/ml) for 1 h and then incubated with nigericin (6.5 µM) for an additional hour [20 (link)]. Then, IL-1β-containing conditioned medium (mIL-1β-CM) were collected to be added to adipocytes. In some experiments, peritoneal macrophages were used instead of BMDM. For human experiments, human peripheral blood mononuclear cells (hPBMC) were purchased from Zenbio and used for hIL-1β-CM preparation. hPBMC were cultured in RPMI 1640 medium with 10% heat-inactivated FBS, 1% penicillin streptomycin, and 2 mM L-glutamine. Adherent monocyte subpopulations were treated for 5 days with 2 ng/ml of human macrophage colony-stimulating factor (Sigma) for macrophage differentiation. These macrophages then were primed with LPS for 1 h and then exposed to nigericin for 1 h to stimulate NLRP3 inflammasome formation and IL-1β secretion.
Okla M., Zaher W., Alfayez M, & Chung S. (2018). Inhibitory Effects of Toll-Like Receptor 4, NLRP3 Inflammasome, and Interleukin-1β on White Adipocyte Browning. Inflammation, 41(2), 626-642.
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