Granulocyte Macrophage Colony Stimulating Factor, Recombinant, Porcine (GM-CSF, Burst Promoting Activity, CMCSF, Colony Stimulating Factor 2, CSF Alpha, Eosinophil Colony Stimulating Factor, Granulocyte Macrophage Colony Stimulating Factor, Molgramostin,
|Molecular Biology||Storage: -20°CShipping: Blue Ice|
The overall structure of GM-CSF is highly compact and globular with a predominantly hydrophobic core, is characterized by a 4-alpha-helix bundle. The helices are arranged in a left-handed anti-parallel fashion, with two overhand connections. Within the connections is a two-stranded anti-parallel beta-sheet. The tertiary structure has a topology similar to that of (pig) growth factor and interferon-beta. Most of the proposed critical regions for receptor binding are located on a continuous surface at one end of the molecule that includes the C terminus.
Granulocyte Macrophage Colony Stimulating Factor is produced in response to a number of inflammatory mediators by mesenchymal cells present in the hemopoietic environment and at peripheral sites of inflammation. Granulocyte Macrophage-CSF is able to stimulate the production of neutrophilic granulocytes, macrophages, and mixed granulocyte-macrophage colonies from bone marrow cells and can stimulate the formation of eosinophil colonies from fetal liver progenitor cells. GM-CSF can also stimulate some functional activities in mature granulocytes and macrophages. GM-CSF receptors shows significant homologies with other receptors for hematopoietic growth factors , including IL2-beta, IL-3 , IL-6 , IL-7 , EPO and the Prolactin receptors
Recombinant Porcine GM-CSF produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 127 amino acids and having a molecular mass of 14381 Dalton.
Porcine GM-CSF is purified by proprietary chromatographic techniques.
Amino acid sequence:
The sequence of the first five N-terminal amino acids was determined and was found to be Ala-Pro-Thr-Arg-Pro.
Dimers and aggregates:
1% as determined by silver-stained SDS-PAGE gel analysis.
Porcine GM-CSF is fully biologically active when compared to standard. The ED50 as determined by the dose-dependent stimulation of the proliferation TF-1 cell line is < 20 ng/ml.
0.1ng/ug (IEU/ug) of GM-CSF .
Protein quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 nm using the absorbency value of 0.977 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 Porcine GM-CSF as a Reference Standard.
Storage and Stability:
Lyophilized powder may be stored at 4°C for short-term only. Reconstitute to nominal volume by adding sterile dH2O or buffer of choice and store at -20°C. 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: E. coli
Purity: 95% as determined by RP-HPLC, anion-exchange FPLC and/or reducing and non-reducing SDS-PAGE Silver Stained gel.
Form: Supplied as a lyophilized powder without additives. It is recommended to reconstitute the lyophilized Porcine GM-CSF in sterile 20mM AcOH (acetic Acid) not less than 100ug/ml, which can then be further diluted to other aqueous solutions.
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.