Certificate of Analysis & Data Sheet

Human Granulocyte Macrophage Colony Stimulating Factor (GM-CSF) 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 . is a single, non-glycosylated, polypeptide chain containing 127 amino acids and having a molecular mass of 14477 Dalton. Recombinant GM-CSF is purified by proprietary chromatographic techniques.

Recombinant Human Granulocyte Macrophage CSF is produced in E.Coli.

Sterile Filtered White lyophilized (freeze-dried) powder.

GM-CSF was lyophilized after extensive dialysis against 2mM sodium phosphate buffer pH= 7.4

It is recommended to reconstitute the lyophilized GM-CSF in sterile DD endotoxin free water less than 1000µg/ml, which can then be further diluted to other aqueous solutions contaning neutral protein as a carrier.

Lyophilized GM-CSF although stable at room temperature for 3 weeks, should be stored desiccated below -180C. Upon reconstitution GM-CSF should be sterilized and stored at 4 C up to 7 days and for future use below -180C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please avoid freeze-thaw cycles.

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
(d) gel filtration on Superdex 75 column.

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.

Less than 1% as determined by gel filtration on Superdex column under non denturative conditions.

PLR's GM-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.

Less than 0.1 ng/µg (IEU/µg) of GM-CSF .

Protein quantitation was carried out by UV spectroscopy at 280 nm using the absorbency value of 0.97 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the DNAman computer analysis program of protein sequences.

This material is offered by PLR for laboratory research