Hgm csf

Manufactured by R&D Systems

Sourced in United States

HGM-CSF is a recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) produced in a mouse myeloma cell line. GM-CSF is a cytokine that stimulates the growth and differentiation of hematopoietic precursor cells, including granulocytes and macrophages.

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Lab products found in correlation

Anti his hrp antibody, by Abcam (1 mentions) Anti human cd14 microbeads, by Miltenyi Biotec (1 mentions) Hil 4, by R&D Systems (1 mentions)

2 protocols using hgm csf

Purification and Bioactivity Evaluation of GM-CSF

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The protein samples were analyzed by 15% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting using anti-His-HRP antibody (Abcam, USA). The concentration of purified GM-CSF was measured using Bradford method.[19 (link)] The biological activity of obtained GM-CSF was determined by evaluation of its proliferative effect on HL-60 cells using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay as described previously.[19 (link)] Briefly, different concentrations (1–100 pg/ml) of expressed and standard hGM-CSF (R and D systems, USA) were added to cells, and after 48 incubation, the viability of cells was evaluated. The percentage of cell survival was plotted against concentrations, and based on the obtained equation, EC50 (the concentration of GM-CSF exhibiting 50% of the maximal proliferative effect) was determined. Specific activity was calculated using the following equation:
Specific activity (IU/μg) = 1/EC50 (pg/ml) × 10⁶ (1 μg = 106 pg)

Taherian E., Mohammadi E., Jahanian-Najafabadi A., Moazen F, & Akbari V. (2019). Cloning, Optimization of Periplasmic Expression and Purification of Recombinant Granulocyte Macrophage-Stimulating Factor in Escherichia coli BL21 (DE3). Advanced Biomedical Research, 8, 71.

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Differentiation of Monocytes to Immature DCs

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Human immature DCs were differentiated from Monocytes as previously described19 (link)21 (link). All experimental protocols were approved by the Institutional Review Boards of Radboud University Medical Center (Netherlands) and Vimercate Hospital (Italy). Peripheral Blood Mononuclear Cells (PBMCs) were isolated from buffy coats (Sanquin) obtained from healthy volunteers after written informed consent and all methods were carried out in accordance with the approved guidelines. Briefly, monocytes were purified from Peripheral Blood Mononuclear Cells (PBMCs) using the anti-human CD14 MicroBeads (Miltenyi Biotech, Calderara di Reno, Italy). Monocytes were differentiated into iMDDCs with hIL-4 and hGM-CSF (both 20 ng/ml; R&D Systems, Minneapolis, USA) for 6 days.

Berzi A., Ordanini S., Joosten B., Trabattoni D., Cambi A., Bernardi A, & Clerici M. (2016). Pseudo-Mannosylated DC-SIGN Ligands as Immunomodulants. Scientific Reports, 6, 35373.

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