/, IL-6 gp130 family, Recombinant Human Cytokines/Human Granulocyte-Colony Stimulating Factor Recombinant

Human Granulocyte-Colony Stimulating Factor Recombinant

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$70.00$4,700.00

SKU: RKP09919 Tags: , , , , , , ,

Description

Accession
P09919
Source
DNA sequence encoding the extracellular domain of Human CSF3 was expressed in E. coli.
Molecular weight
Recombinant Human CSF3 is a protein consisting of amino acid residues [Ala 30-Pro204] ,and migrates as an approximately 19 kDa protein on 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 < 0.1 ng/ml, corresponding to a specific activity of ≥ 1x 10 8 units/mg.
Endotoxin
Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than 0.1 ng/µg(1EU/µg).
Presentation
Recombinant CSF3 was lyophilized from 0.2 µm filtered PBS solution pH7.4
Sequence
ATPLGPASSL PQSFLLKCLE QVRKIQGDGA ALQEKLVSEC ATYKLCHPEE LVLLGHSLGI PWAPLSSCPS QALQLAGCLS QLHSGLFLYQ GLLQALEGIS PELGPTLDTL QLDVADFATT IWQQMEELGM APALQPTQGA MPAFASAFQR RAGGVLVASH LQSFLEVSYR
Reconstitution
A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than 0.1 mg/mL. This solution can then be diluted into other buffers.
Storage
The lyophilized protein is stable for at least 2 years from date of receipt at -20° C.
Usage
This product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.
Interactor
P40223
Molecular function
Molecular function

Methods

Bioactivity Assay

  • The proliferation assay of NFS-60 cells in response to the presence of hG-CSF measured by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] method was used to test the bioactivity of expressed hG-CSF in plants.
  • NFS-60 cells were cultured with medium'>RPMI-1640 medium supplemented with 10% fatal bovine serum (FBS) and commercial recombinant hG-CSF (2 ng/ml) and kept under 5% CO2 at 37 °C in a humidified condition.
  • For MTT test, total soluble protein extracted from leaves and seeds (see Protein extraction and immunoblot above) were used for analysis and NFS-60 cells (1 × 104 per well) were cultured in a 96 well plate and treated with the medium medium'>(RPMI-1640 medium supplemented with 10% FBS) containing the following samples: commercial recombinant hG-CSF (1 ng); total soluble protein extracted from leaf (TSP-L) and seed (TSP-S) of transgenic plant harboring the Construct USH containing 1…
  • CD34 Cord blood (CB) samples were obtained from the Hannover Medical School following written consent of the donors as approved by the Hannover Medical School local ethics committee.
  • Total nucleated cells were isolated by a Ficoll gradient followed by enrichment for CD34+ cells employing MACS purification .
  • Isolated cells were frozen until further usage.
  • CB-CD34+ cells were cultured in medium'>StemSpan medium supplemented with 1% penicillin/streptomycin and one of four cytokine combinations using hSCF, hTHPO, hFLT3-L, hGCSF, hIGFBP2, hAngptl5, hFGF-1, and hIL6 .
  • The StemRegenin compound was used at a concentration of 1 µmol/l as described.
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