Human beta Nerve Growth Factor, CHO (Recombinant)

Recombinant Human beta-Nerve Growth Factor (NGF-b)

Source: CHO cells

Background: Nerve Growth Factor is the first of a series of neurotrophic factors that were found to influence the growth and differentiation of sympathetic and sensory neurons. It is comprised of alpha, beta, and gamma subunits. The beta subunit is responsible for its growth stimulating activity. The synthesis of NGF in astrocytes is enhanced by various other cytokines: IL-1 , TNF-alpha , PDGF and TGF-beta .

Description: Recombinant Human Nerve Growth Factor beta produced in CHO is a homodimer, glycosylated, polypeptide chain containing 2 identical 119 amino acids and having a molecular mass of 16,950 Dalton. Human Nerve Growth Factor is purified by proprietary chromatographic techniques.

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

Formulation: NGF-beta was lyophilized from a 0.2 m filtered solution in 30mM sodium acetate pH=5.5 containing 1% HSA.

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

Stability: Lyophilized NGF-b although stable at room temperature for 3 weeks, should be stored desiccated below -18 C. Upon reconstitution NGF-b 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 Ser-Ser-Ser-His-Pro.

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

Biological Activity: beta-Nerve Growth Factor is fully biologically active when compared to standards. The ED50, calculated by its ability to stimulate chick E9 DRG neurite outgrowth was found to be < 1.0 ng/ml, corresponding to a specific activity of > 1 x 106 units/mg.

Endotoxin: Less than 0.1 ng/ g (IEU/ g) of Recombinant beta-Nerve Growth Factor.

Protein content: Protein quantitation was carried out by two independent methods: 1. UV spectroscopy at 280 nm using the absorbency value of 1.1945 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 Recombinant beta-Nerve Growth Factor as a Reference Standard.