Human Glial-Derived Neurotrophic Factor (Recombinant)

Recombinant Human Glial-derived Neurotrophic Factor (GDNF)

Source: Baculovirus

Background: This factor was found initially in conditioned medium of the rat B49 glial cell line.GDNF is a disulfide-bonded homodimeric glycosylated protein of 134 amino acids .The proteins contain seven conserved cysteine residues in the same relative spacing found in all members of the TGF-beta superfamily of proteins. The human GDNF gene has been mapped to chromosome 5p13.1-p13.3 . Related factors are Neurturin , Persephin , and Artemin . Collectively they are being referred to as GDNF family ligands .

Description: Recombinant Human Glial-derived Neurotrophic Factor (GDNF) produced in E.Coli is a homodimer, non-glycosylated, polypeptide chain containing 2 x 135 amino acids and having a total molecular mass of 30,360 Dalton. Recombinant GDNF is purified by proprietary chromatographic techniques.

Physical Appearance: Sterile Filtered White lyophilized (freeze-dried) powder.

Formulation: Recombinant GDNF was lyophilized after dialysis against 10 mM sodium citrate and 150mM NaCl.

Solubility: It is recommended to reconstitute the lyophilized GDNF in sterile 18MO-cm H2O not less than 100 g/ml, which can then be further diluted to other aqueous solutions.

Stability: Lyophilized GDNF although stable at room temperature for 3 weeks, should be stored desiccated below -18 C. Upon reconstitution GDNF 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 avoid freeze-thaw cycles.

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.

Amino-Acid Sequence: The sequence of the first five N-terminal amino acids was determined and was found to be Met-Ser-Pro-Asp-Lys.

Dimers and aggregates: Less than 1% as determined by silver-stained SDS-PAGE gel analysis.

Biological Activity: Glial-Derived Neurotrophic Factor is fully biologically active when compared to standard. The ED50, calculated by the dose-dependant dopamin uptake in rat mesencephalic cultures was found to be 5-10 ng/ml.

Endotoxin: Less than 0.1 ng/ g (IEU/ g) of Recombinant Glial-derived Neurotrophic Factor.

Protein content: Protein quantitation was carried out by two independent methods: 1. UV spectroscopy at 280 nm . 2. Analysis by RP-HPLC, using a calibrated solution of Recombinant Glial-derived Neurotrophic Factor as a Reference Standard.

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