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Human GM-CSF, amino acids Ala18- Glu144 (Accession# NM_000758) was expressed in E. coli.
Molecular Mass:
The 127 amino acid N-terminal methionylated recombinant protein has a predicted molecular mass of 14477.5 Da. The DTT-reduced protein migrates at approximately 14 kDa and the non-reduced protein migrates with slightly greater mobility by SDS-PAGE.
Purity:
>98%, as determined by Coomassie stained SDS-PAGE.
Formulation:
0.22µm filtered protein solution is in PBS.
Endotoxin Level:
Less than 0.01ng per µg cytokine as determined by the LAL method.
Concentration:
10-100µg sizes are bottled at 200µg/mL, 500µg sizes and larger are bottled at the concentration indicated on the vial.
Storage & Handling:
Unopened vial can be stored at 4°C for three months or -20°C to -70°C for six months. For maximum results, quick spin vial prior to opening. Stock solutions should be prepared at no less than 10µg/mL in buffer containing carrier protein such as 1% BSA or HSA or 10% FBS. For long term-storage, aliquot into polypropylene vials and store in a manual defrost freezer. Avoid repeated freeze/thaw cycles.
Activity:
The ED50 is 0.10- 0.30 ng/ml, corresponding to a specific activity of 1 - 0.33 X107 units/mg, as determined by a dose dependent TF-1 cell proliferation.
GM-CSF plays a key role in signaling emergency hemopoiesis (predominantly myelopoiesis) in response to infection, including the production of granulocytes and macrophages in the bone marrow and their maintenance, survival, and functional activation at sites of injury or insult. The receptor for GM-CSF is a heterodimer that comprises a major binding subunit (GMRα) and a major signaling subunit (βc). The receptor subunits are always coexpressed on the surface of leukocytes, with βc being expressed at lower levels than GMRα. Certain nonhemopoietic cell types have also been reported to express the GM-CSF receptor and to respond to GM-CSF stimulation in vitro, although the in vivo significance of these observations remains uncertain.
T cells, macrophages, fibroblasts, endothelial cells, mast cells.
Function:
Synergistic with IL-1, IL-3, G-CSF; E21R competitive antagonist for receptor binding; stored in ECM with heparan sulfate proteoglycans
Ligand Receptor:
Heterodimer GM-CSFR α subunit (CDw116); β-subunit (CDw131) in common
Interaction:
Granulocyte/macrophage/erythroid/megakaryocytic progenitors, myeloblasts, monoblasts, dendritic cells, T cells.
Bioactivity/Activities:
Granulocyte-macrophage colony-stimulating factor (GM-CSF) and the related cytokines interleukin (IL)–3 and IL-5 regulate the production and functional activation of hemopoietic cells, with GM-CSF acting on monocyte/macrophages and all granulocytes. GM-C
Antigen References:
1. Hercus TR, et al. 2009 Blood 114:1289-1298. 2. Hayashida K, et al. 1990 Proc Natl Acad Sci USA. 87:9655-9659. 3. Walker F, et al. 1985 EMBO J 4:933-939. 4. Kitamura T, et al. 1989 J. Cell Physiol. 140:323.
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