- Description
- Specifications
Catalog #: CYT-221
Description:
Granulocyte Macrophage Colony Stimulating Factor Human Recombinant produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 127 amino acids and having a molecular mass of 14477 Dalton. GM-CSF is purified by proprietary chromatographic techniques.
Synonyms:
CSF-2, MGI-1GM, GM-CSF, Pluripoietin-alpha, Molgramostin, Sargramostim, MGC131935, MGC138897.
Source:
Escherichia Coli.
Amino Acid Sequence:
The sequence of the first five N-terminal amino acids was determined and was found to be Ala-Pro-Ala-Arg-Ser.
N-terminal methionine has been completely removed enzymatically.
Purity:
Greater than 98.0% as determined by1. Analysis by RP-HPLC.
2. Analysis by SDS-PAGE.
Solubility:
It is recommended to reconstitute the lyophilized Granulocyte Macrophage Colony Stimulating Factor in sterile 18MΩ–cm H2O not less than 100 µg/ml, which can then be further diluted to other aqueous solutions.
Formulation:
GM-CSF was lyophilized after extensive dialysis against 2mM sodium phosphate buffer pH= 7.4 ±0.1.
Stability:
Lyophilized Granulocyte Macrophage Colony Stimulating Factor although stable at room temperature for 3 weeks, should be stored desiccated below -18 °C. Upon reconstitution GMCSF should be stored at 4 °C between 2-7 days and for future use below -18 °C.
For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please prevent freeze-thaw cycles.
Activity:
The ED50 as determined by the dose-dependant stimulation of the proliferation of human TF-1 cells (human erythroleukemic indicator cell line) is < 0.1 ng/ml, corresponding to a Specific Activity of 11,100,000 IU/mg.
Physical Appearance:
Sterile Filtered White lyophilized (freeze-dried) powder.
Protein Content:
GM-CSF quantitation was carried out by two independent methods
1. UV spectroscopy at 280 nm using the absorbency value of 0.963 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the PC GEN computer analysis program of protein sequences (IntelliGenetics).
2. Analysis by RP-HPLC, using a standard solution of GM-CSF as a Reference Standard.
Usage:
Denovo Biotechnology's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
References:
1. Title: Disease activity in systemic lupus erythematosus is associated with an altered expression of low-affinity Fc gamma receptors and costimulatory molecules on dendritic cells.
Publication: Immunology. 2009 Nov;128(3):334-41. PMID: 2770681.
Link: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2770681/
Applications: IL-4 and GM-CSF were used to obtain monocyte-derived Dendritic cells(DC) from Human peripheral blood mononuclear cells (PBMCs).
2. Title: Generation of Novel Bone Forming Cells (Monoosteophils) from the Cathelicidin-Derived Peptide LL-37 Treated Monocytes.
Publication: PLoS One. 2010; 5(11): e13985. Published online 2010 November 15.
Link: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2981577/?tool=pmcentrez
Applications: GM-CSF was used to differentiate monocytes into macrophage and to Monocyte-derived dendtritic cells (DCs) from Peripheral blood mononuclear cells (PBMCs).
Osteoclasts, which are derived from monocytes by the action of macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor ?B ligand (RANKL). Osteoclast were differentiated from monocytes in the presence of RANKL and M-CSF (both at 25 ng/mL).
And also for cytokine analysis.
3.Title:AIMP1/p43 protein induces the maturation of bone marrow-derived dendritic cells with T helper type 1-polarizing ability.
Publication: J Immunol. 2008 Mar 1;180(5):2894-902. PMID: 18292511.
Link: http://www.ncbi.nlm.nih.gov/pubmed?term=18292511
Applications: Generation of bone marrow DCs.
4. Title:Sphingosine kinase regulates the rate of endothelial progenitor cell differentiation.
Publication: Blood. 2009 Feb 26;113(9):2108-17. Epub 2008 Dec 24. PMID: 19109558. PMID: 18292511.
Link: http://www.ncbi.nlm.nih.gov/pubmed?term=19109558
Applications: BM cells at 10,000 were plated in 0.3% agar culture medium containing GM-CSF (100 ng/ml, was used with other cytokines for mouse progenitor cell isolation, culture and colony forming assay.
5.Title:Defective IL-10 production in severe phenotypes of Crohn's disease
Publication:Published online before print February 23, 2009, doi: 10.1189/jlb.1108698 May 2009 Journal of Leukocyte Biology vol. 85 no. 5 896-903
Link:http://www.jleukbio.org/content/85/5/896.full
6.Title:Adenosine deaminase potentiates the generation of effector, memory, and regulatory CD4+ T cells .
Publication:Published online before print October 19, 2010, doi: 10.1189/jlb.1009696 January 2011 Journal of Leukocyte Biology vol. 89 no. 1 127-136
Link:http://www.jleukbio.org/content/89/1/127.full
7.Title:Glutamate Released by Dendritic Cells as a Novel Modulator of T Cell Activation.
Publication:
Link:http://journal.9med.net/qikan/article.php?id=367793
8.Title:Airway Epithelial Cells Regulate the Functional Phenotype of Locally Differentiating Dendritic Cells: Implications for the Pathogenesis of Infectious and Allergic Airway Disease1.
Publication: The Journal of Immunology January 1, 2009 vol. 182 no. 1 72-83
Link:http://www.jimmunol.org/content/182/1/72.full
9.Title:A New Multi-clade DNA Prime/Recombinant MVA Boost Vaccine Induces Broad and High Levels of HIV-1-specific CD8+ T-cell and Humoral Responses in Mice.
Publication:Received 31 October 2006; Accepted 14 May 2007; Published online 19 June 2007.
Link:http://www.nature.com/mt/journal/v15/n9/full/6300235a.html
