The sequence of the first five N-terminal amino acids of GCSF was determined and was found to be Met-Thr-Pro-Leu-Gly.
The ED50, calculated by the dose-dependant proliferation of murine NFS-60 indicator cells (measured by 3H-thymidine uptake) is < 0.1 ng/ml, corresponding to a Specific Activity of 100,000,000 IU/mg.
GCSF was lyophilized after extensive dialysis against 10mM sodium acetate buffer pH= 4.
Sterile Filtered White lyophilized (freeze-dried) powder.
GCSF quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 nm using the absorbency value of 0.815 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 GCSF as a Reference Standard.
Greater than 98.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Analysis by SDS-PAGE.
It is recommended to reconstitute the lyophilized GCSF in sterile 20mM AcOH not less than 100ug/ml, which can then be further diluted to other aqueous solutions.
Lyophilized GCSF although stable at room temperature for 3 weeks, should be stored desiccated below -18oC. Upon reconstitution GCSF should be stored at 4oC between 2-7 days and for future use below -18oC.
For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please prevent freeze-thaw cycles.
CSF-3, MGI-1G, GM-CSF beta, Pluripoietin, Filgrastim, Lenograstim, G-CSF, MGC45931, GCSF.
