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Description |
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.
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Source |
Escherichia Coli. |
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Physical Appearance: |
Sterile Filtered White lyophilized (freeze-dried) powder.
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Formulation: |
The protein was lyophilized after extensive dialysis against 2mM sodium phosphate buffer pH= 7.4±0.1.
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Solubility: |
It is recommended to reconstitute the lyophilized rHuGM-CSF in sterile 18MΩ-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
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Stability: |
Lyophilized rHuGM-CSF although stable at room temperature for 3 weeks, should be stored desiccated below -18 °C. Upon reconstitution rHuGM-CSF 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.
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Purity: |
Greater than 98.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel. |
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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.
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Dimers and aggregates: |
Less than 1% as determined by silver stained SDS-PAGE gel analysis.
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Biological Activity: |
ProSpec's rHuGM-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 less then 0.1 ng/ml, corresponding to a Specific Activity of 11.1x106 IU/mg.
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Endotoxin: |
Less than 0.1 ng/µg (IEU/µg) of rHuGM-CSF.
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Protein content: |
Protein quantitation was carried out by two independent methods:
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). Analysis by RP-HPLC, using a standard solution of GM-CSF as a Reference Standard. |
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Usage |
This material is for research, laboratory or further manufacturing purposes. |