Mouse Granulocyte Colony Stimulating Factor Recombinant

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Mouse Granulocyte Colony Stimulating Factor Recombinant

$70.00$4,700.00


accession P09920


Source Optimized DNA sequence encoding mouse Granulocyte Colony Stimulating Factor mature chain was expressed in Escherichia Coli.
Molecular weight Native mouse G-CSF is generated by the proteolytic removal of the signal peptide and propeptide, the molecule has a calculated molecular mass of approximately 19kDa. Recombinant GCSF is a monomer protein consisting of 179 amino acid residue subunits, and migrates as an approximately 19kDa protein under non-reducing and reducing conditions in SDS-PAGE.
Purity >95%, as determined by SDS-PAGE and HPLC
Biological Activity The ED(50) was determined by the dose-dependent proliferation of murine M-NFS-60 cells is <.05 ng/ml, corresponding to a specific activity of > x units/mg.
Protein Sequence MAQLSAQRRM KLMALQLLLW QSALWSGREA VPLVTVSALP PSLPLPRSFL LKSLEQVRKI QASGSVLLEQ LCATYKLCHP EELVLLGHSL GIPKASLSGC SSQALQQTQC LSQLHSGLCL YQGLLQALSG ISPALAPTLD LLQLDVANFA TTIWQQMENL GVAPTVQPTQ SAMPAFTSAF QRRAGGVLAI SYLQGFLETA RLALHHLA
Endotoxin Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than.1 ng/µg(1EU/µg).
Presentation IRecombinant mouse G-CSF was lyophilized from a.2 μm filtered PBS solution.
Reconstitution A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than.1 mg/mL. This solution can then be diluted into other buffers.
Storage The lyophilized protein is stable for at least years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at° -° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
Usage This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.


Interactor Q99062 CSF3R_HUMAN
Molecular function Cytokine
Molecular function Growth-factor

Methods

Inhibition of neutrophil differentiation by IRF4.

G-CSF
  • Wright-Giemsa staining of 32Dcl.3 cells transduced with empty MSCV-puro, MSCV-IRF4FLAG-puro or MSCV-IRF8FLAG-puro and cultured in the presence of IL-3 (upper panels) or G-CSF (for 7 days, lower panels).

G-CSF production by various mouse tumor models.

recombinant G-CSF
  • Sera G-CSF levels determined from nontumor-bearing mice (n = 10), mice treated with recombinant G-CSF protein (10 µg daily for 5 consecutive days) (n = 8) or mice bearing 4T1 cells (n = 9) or AT-3 tumor cells pooled from two separate experiments (n = 9; overall tumor volume ≤2cm3).

Generation of conditional TAK1-deficient mice.

10 ng/ml G-CSF
  • Colony formation by BM cells from Map3k7

Bone marrow transduction

  • Retroviral transduction of bone marrow was done as previously described (6/ml) were prestimulated for 48 hours in IDMEM containing 10% Premium FBS , 2% Penicillin/Streptomycin, and 100 ng/ml each of recombinant murine TPO, G-CSF, and SCF .
  • For primary transduction, cells were resuspended in appropriate retroviral supernatant containing growth factors and seeded onto fibronectin coated wells ((

Generation of MDSCs and DCs from BM

  • MDSCs were generated from BM of naïve wt or EGFP-LysM-Tg BALB/c mice.
  • Femurs and tibias were collected under aseptic condition, and BM was flushed out with sterile PBS.
  • After red blood cell lysis, BM cells were counted (the number of cells was usually 3–4×107 per mouse), and seeded in Petri dishes at a density of 5×105 cells per ml of Dulbecco’s Modified Eagle Medium (DMEM;) containing 10% fetal bovine serum (FBS) .
  • In preliminary dose-finding experiments the BM cells were cultured for 3 to 7 days in the presence of varying doses of recombinant murine granulocyte macrophage colony stimulating factor (rmGM-CSF, , ) and recombinant murine interleukin-6 (rmIL-6), or with a combination of rmGM-CSF, rmIL-6, and recombinant murine granulocyte colony stimulating factor (rmG-CSF).
  • On the basis of phenotypic and functional characteristics, the optimal protocol for BM-MDSC
  • MDSCs were generated from BM of naïve wt or EGFP-LysM-Tg BALB/c mice.
  • Femurs and tibias were collected under aseptic condition, and BM was flushed out with sterile PBS.
  • After red blood cell lysis, BM cells were counted (the number of cells was usually 3–4×107 per mouse), and seeded in Petri dishes at a density of 5×105 cells per ml of Dulbecco’s Modified Eagle Medium (DMEM;) containing 10% fetal bovine serum (FBS) .
  • In preliminary dose-finding experiments the BM cells were cultured for 3 to 7 days in the presence of varying doses of recombinant murine granulocyte macrophage colony stimulating factor (rmGM-CSF, , ) and recombinant murine interleukin-6 (rmIL-6), or with a combination of rmGM-CSF, rmIL-6, and recombinant murine granulocyte colony stimulating factor (rmG-CSF).
  • On the basis of phenotypic and functional characteristics, the optimal protocol for BM-MDSC generation was found to be a 3-day culture of BM cells in the presence of rmGM-CSF, rmIL-6, and rmG-CSF (10 ng/ml each).

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Phenotype and morphology of myeloid-derived suppressor cell (MDSC)-like cells generated in vitro from murine bone marrow (BM) in comparison with synovial fluid (SF) cells.

G-CSF
  • BM cells were cultured in the presence of GM-CSF, IL-6, and G-CSF (10 ng/ml each).

Cell culture and retroviral transduction

  • Kasumi-1, 293T and U937 cells were cultured as previously described. G-CSF (granulocyte colony-stimulating factor)-mobilized peripheral blood CD34+ cells were cultured in Iscove's medium'>modified medium'>Dulbecco's medium supplemented with 20% fetal calf serum, 20 ng/ml Flt-3 l, 20 ng/ml GM-CSF, 20 ng/ml stem cell factor (SCF), 20 ng/ml thrombopoietin, 20 ng/ml interleukin (IL)-6, 10 ng/ml IL-3 (all cytokines were obtained, , ), 100 U/ml penicillin/streptomycin and 2 mM L-glutamine.
  • Retroviral transduction and long-term cultivation were performed as previously described. IMR-90 cells (ATCC-CCL-186) were cultured as suggested by the supplier.