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Recombinant Human Granulocyte Macrophage Colony
Stimulating Factor GM CSF
(mGMP-rHuGM-CSF)
Description:
mGMP Recombinant Human GM-CSF produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 127 amino acids and having a molecular mass of 14477 Dalton. rHuGM-CSF is purified by proprietary chromatographic techniques.
Source:
Escherichia Coli.
Physical Appearance:
Sterile Filtered White lyophilized (freeze-dried) powder.
Formulation & Packaging:
The protein was lyophilized after extensive dialysis against 2mM sodium phosphate buffer pH= 7.4�0.1.
Solubility:
The lyophilized rHuGM-CSF is very soluble in water and most aqueous buffers below and above the isoelectric point (pI=4.95).
Stability:
Lyophilized mGMPrHuGM-CSF although stable at room temperature, should be stored desiccated below 0�C. Reconstituted rHuGM-CSF is best stored refrigerated at 4�C.
Purity:
Greater than 99.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained.
(Limit of acceptance:�98.0%. No more than 2% total impurities; no single impurity greater than 1%)
Amino Acid Composition:
In total agreement with the expected amino acid composition of native
human GM-CSF.
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, conforming the sequence of native human GM-CSF. N-terminal methionine has been completely removed enzymatically.
Dimers and aggregates:
Less than 1% as determined by silver stained SDS-PAGE gel analysis.
Biological Activity:
mGMPrHuGM-CSF is fully biologically active when compared to standard.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 9x106 IU/mg or 2 800 000 IU/ vial of 300 ug
Endotoxin:
Less than 0.1 ng/�g (IEU/�g) of mGMPrHuGM-CSF.
Protein content:
Protein 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 GENE
computer analysis program of protein sequences (IntelliGenetics).
2. Analysis by RP-HPLC, using a standard solution of GM-CSF as a Reference Standard.
Usage:
This material is offered by Gentaur for research, laboratory or further manufacturing purposes adn DC culture
Cell lines:
Hematopoietic cell lines K562, human erythroleukemia cell line (HEL), HL60, TF-1, Jurkat, Daudi, CEM, and ML1 (ATCC, Manassas, VA, USA) were cultured in the presence of 10% fetal bovine serum (FBS) (Euroclone, Milano, Italy) in Iscove's modified Dulbecco's medium (IMDM) or RPMI (Euroclone, Milano Italy) and 5% CO2 in a humidified atmosphere of 5% O2 in air at 37�C. TF-1 required the addition of 10 ng/ml of human granulocyte-macrophage colony-stimulating factor (GM-CSF, Leukomax; Schering-Plough, Basel, Switzerland).
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| GM CSF | mGMP-rHuGM-CSF | 300�g | 385 Euro |
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