Detection of G‑CSF in Human Macrophages by Flow Cytometry.
Human macrophages treated with LPS, Recombinant Human TNF‑ alpha (Catalog # 210-TA), and Recombinant Human IL‑ beta (Catalog # 201-LB) were stained with Mouse Anti-Human G‑CSF PE‑conjugated Monoclonal Antibody (Catalog # IC2141P, filled histogram) or isotype control antibody (Catalog # IC002P, open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Preparation and Storage
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
12 months from date of receipt, 2 to 8 °C as supplied.
G-CSF is a pleiotropic cytokine best known for its specific effects on the proliferation, differentiation, and activation of hematopoietic cells of the neutrophilic granulocyte lineage. It is produced mainly by monocytes and macrophages upon activation by endotoxin, TNF-alpha and IFN-gamma. Other cell types including fibroblasts, endothelial cells, astrocytes and bone marrow stromal cells can also secrete G-CSF after LPS, IL-1, or TNF-alpha activation. In addition, various carcinoma cell lines and myeloblastic leukemia cells can express G-CSF constitutively.
In humans, two distinct cDNA clones for G-CSF, encoding 207 and 204 amino acid precursor proteins, have been isolated. Both proteins have a 30 amino acid signal peptide and have identical amino acid sequences except for a three amino acid insertion (deletion) at the 35th amino acid residue from the N-terminus of the mature protein. Human G-CSF is 73% identical at the amino acid level to murine G-CSF and the two proteins show species cross-reactivity.
In vitro, G-CSF stimulates growth, differentiation and functions of cells from the neutrophil lineage. It also has blast cell growth factor activity and can synergize with IL-3 to shorten the Go period of early hematopoietic progenitors. Consistent with its in vitro functions, G-CSF has been found to play important roles in defense against infection, in inflammation and repair, and in the maintenance of steady state hematopoiesis.
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