Granulocyte Colony Stimulating Factor, Recombinant, Human (G-CSF)
A glycoprotein of MW 20kD containing internal disulfide bonds. It induces the survival, proliferation, and differentiation of neutrophilic granulocyte precursor cells and functionally activates mature blood neutrophils. Among the family of colony-stimulating factors, G-CSF is the most potent inducer of terminal differentiation to granulocytes and macrophages of leukemic myeloid cell lines. The synthesis of G-CSF can be induced by bacterial endotoxins, TNF, Interleukin-1 and GM-CSF. Prostaglandin E2 inhibits the synthesis of G-CSF. In epithelial, endothelial, and fibroblastic cells secretion of G-CSF is induced by Interleukin-17.
Recombinant Human Granulocyte Colony Stimulating Factor is a glycosylated, polypeptide chain containing 174 amino acids and having a molecular mass of 20kD.
Biological Activity
G-CSF is fully biologically active when compared to standard. The ED50, calculated by the dose-dependent proliferation of murine NFS-60 indicator cells (measured by 3H-thymidine uptake) is less then 0.1 ng/ml, corresponding to a Specific Activity of 1.27 x 10e8 IU/mg.
G-CSF protein quantitation was carried out by two independent methods
1. UV spectroscopy at 280nm using the absorbency value of 0.815 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics). 2. Analysis by RP-HPLC, using a calibrated solution of Recombinant Human Granulocyte Colony Stimulating Factor as a Reference Standard.
Sequence
The sequence of the first five N-terminal amino acids was determined and was found to be Thr-Pro-Leu-Gly-Pro.
Storage and Stability
Lyophilized powder may be stored at 4°C for short-term only. Reconstituted product is stable for 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Source
Recombinant, CHO cells
Purity
≥ 98% by RP-HPLC, FPLC, or reducing/non-reducing SDS-PAGE Silver Stain. Chromatographically purified.
Form
Supplied as a lyophilized powder in 10mM HCl, pH 6.5, Tween 20, mannitol, L-arginine, phenylalanine, methionine.
Important Note
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
