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Amino Acid Sequence: TPLGPASSLP QSFLLKCLEQ VRKIQGDGAA LQEKLCA TYKLCHPEEL VLLGHSLGIP WAPLSSCPSQ ALQLAGCLSQ LHSGLFLYQG LLQALEGISP ELGPTLDTLQ LDVADFATTI WQQMEELGMA PALQPTQGAM PAFASAFQRR AGGVLVASHL QSFLEVSYRV LRHLAQP
Background: G-CSF is secreted by monocytes, macrophages, and neutrophils after cell activation. It is produced also by stromal cells, fibroblasts, and endothelial cells, epithelial carcinomas, acute myeloid leukemia cells and various tumor cell lines. The synthesis of G-CSF can be induced by bacterial endotoxins, TNF, IL1 and GM-CSF. Comparison of the primary sequence of G-CSF with those of the two other colony stimulating factors, GM-CSF and M-CSF, shows that the three factors are not related to each other. Murine and human G-CSF show a sequence homology of approximately 70% at the DNA level and of 72% at the protein level. The G-CSF receptor, CD114, is expressed on all cells of the neutrophils and granulocytes lineage, as well as placenta cells, endothelial cells and various carcinoma cell lines .Human G-CSF is active in murine cells and vice versa. G-CSF stimulates the proliferation and differentiation of hematopoietic progenitor cells committed to the neutrophils and granulocytes lineage in a dose-dependent manner. G-CSF synergises with some other cytokines, including GM-CSF and IL-4, for example, GM-CSF and G-CSF are required to develop neutrophilic colonies in vitro. The concerted action of G-CSF and Epo is required to support the growth of mixed colonies of the early erythroid progenitors. A combination of IL-4 with G-CSF has been shown to lead to synergistic suppression of the growth of some human leukemic cell lines.
Biological Activity: The ED50 was determined by the dose-dependent proliferation of murine M-NFS-60 cells is ≤ 0.1 ng/ml, corresponding to a specific activity of≥ 1 x 108units/mg.
Description: Recombinant GCSF is a disulfide-linked monomer protein consisting of 175 amino acid residues, and migrates as an approximately 19 kDa protein under non-reducing and reducing conditions in SDS-PAGE. Optimized DNA sequence encoding Human G-CSF mature chain was expressed in E. coli.
Endotoxin Level: <0.1 ng/µg (1 EU/µg), using the LAL gel clot method.
Format: lyophilized protein
Formulation: Lyophilized from a 0.2 µm filtered PBS solution pH 7.0.
Genbank: P09919
Purity: ≥98% by SDS-PAGE and HPLC
Reconstitution: Reconstitute at 0.1-1.0 mg/ml in distilled water. This solution can then be diluted into other buffers. To maximize product collection from vial surface, vortex briefly and then spin down to recollect the liquid.
Storage Stability: The lyophilized protein is stable for at least 2 years from date of receipt when stored at -20°C. Upon reconstitution, store in working aliquots at +4°C for up to one month, or at -20°C for up to six months, in the presence of a carrier protein. Avoid repeated freeze/thaw cycles.
Uniprot: P09919
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Donahue, R., et al. Blood September 17, 2009 vol. 114 no. 12 2530-2541.
2. Worm, J., et al. Nucl. Acids Res. (2009) 37 (17): 5784-5792.
3. Diederich, K., et al. The Journal of Neuroscience, 16 September 2009, 29(37): 11572-11581,
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