Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Human Granulocyte Colony Stimulating Factor (hG-CSF) #8930

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Source

Recombinant human G-CSF (hG-CSF) Thr31-Pro204 (Accession #NP_757373) was expressed in human 293 cells at Cell Signaling Technology.

Molecular Characterization

Recombinant hG-CSF contains no "tags" and the nonglycosylated protein has a calculated MW of 18,986. DTT-reduced and non-reduced protein migrate as 18 kDa polypeptides. The expected amino-terminal TPLGP of recombinant hG-CSF was verified by amino acid sequencing.

Purity

>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hG-CSF. All lots are greater than 98% pure.

Bioactivity

The bioactivity of recombinant hG-CSF was determined in a M-NFS-60 cell proliferation assay. The ED50 of each lot is between 20-150 pg/ml.

Coomassie Gel

Coomassie Gel

The purity of recombinant hG-CSF was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hG-CSF and staining overnight with Coomassie Blue.

Bioactivity

Bioactivity

The proliferation of M-NFS-60 cells treated with increasing concentrations of hG-CSF was assessed. After 72 hour treatment with hG-CSF, cells were incubated with a tetrazolium salt and the OD450 - OD650 was determined.

Western Blotting

Western Blotting

Western blot analysis of extracts from M-NSF-60 cells untreated or treated with hG-CSF for 15 minutes, using Phospho-Stat3 (Tyr705) (D3A7) Rabbit mAb #9145 (upper) or Stat3 Antibody #9132 (lower).


Endotoxin

Less than 0.01 ng endotoxin/1μg hG-CSF.

Formulation

With carrier: Lyophilized from a 0.22 μm filtered solution of 40 mM phosphate pH 4.0 containing 250 mM NaCl and 20 μg BSA per 1 μg hG-CSF. Carrier free: Lyophilized from a 0.22 μm filtered solution of 40 mM phosphate pH 4.0 containing 250 mM NaCl.

Background

G-CSF is a hematopoietic cytokine essential for neutrophil development, survival, and egress from bone marrow (1-4). Macrophages and monocytes are the predominant producers of G-CSF (3) and endothelial cells, fibroblasts and neuronal cells can produce G-CSF in response to inflammatory stimuli (3). G-CSF inhibits apoptosis in neutrophils and neurons (4,5). G-CSF stimulates proliferation and differentiation of neuronal progenitor cells (5). G-CSF binding to G-CSFR induces receptor dimerization and activation of Jak1/2 tyrosine phosphorylation (3,6). Signaling is through Stat3, ERK, p38, and Akt (5,6). Absence of functional G-CSF or its receptor in humans and mice causes neutropenia (7,8).

  1. Furze, R.C. and Rankin, S.M. (2008) Immunology 125, 281-8.
  2. Demetri, G.D. and Griffin, J.D. (1991) Blood 78, 2791-808.
  3. Srinivasa, S.P. and Doshi, P.D. (2002) Leukemia 16, 244-53.
  4. van Raam, B.J. et al. (2008) Blood 112, 2046-54.
  5. Schneider, A. et al. (2005) J Clin Invest 115, 2083-98.
  6. Nicholson, S.E. et al. (1994) Proc Natl Acad Sci U S A 91, 2985-8.
  7. Lieschke, G.J. et al. (1994) Blood 84, 1737-46.
  8. Dong, F. et al. (1994) Proc Natl Acad Sci U S A 91, 4480-4.

Application References

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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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