Human Ciliary Neurotrophic Factor (Recombinant)

Recombinant Human Ciliary Neurotrophic Factor (CNTF)

Source: E.Coli

Background: A neurotrophic factor that promotes the survival of various neuronal cell types and may play an important role in the injury response in the nervous system. CNTF, like FGF acidic , FGF basic , and PD-ECGF (platelet-derived endothelial cell growth factor ), does not possess a signal sequence that would allow secretion of the factor by classical secretion pathways (endoplasmatic reticulum/Golgi system). The mechanism underlying the release of CNTF is unknown.

Description: Recombinant Human CNTF produced in E.Coli is a single, non-glycosylated polypeptide chain containing 199 amino acids and having a molecular mass of 22706 Dalton. Recombinant CNTF is purified by proprietary chromatographic techniques.

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

Formulation: Lyophilized from a concentrated (1mg/ml) solution in water containing 5mM sodium Phosphate buffer pH=7.5 and 5mM sodium chloride.

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

Stability: Lyophilized CNTF although stable at room temperature for 3 weeks, should be stored desiccated below -18 C. Upon reconstitution CNTF 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 gel.

Amino acid sequence: The sequence of the first five N-terminal amino acids was determined and was found to be Met-Ala-Phe-Thr-Glu.

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

Biological Activity: Recombinant Human Ciliary Neurotrophic Factor is fully biologically active when compared to standard. The ED50 as determined by the dose-dependant stimulation of TF-1 cells is less then 2 ng/ml, corresponding to a Specific Activity of 5 x 10 IU/mg.

Endotoxin: Less than 0.1 ng/ g (IEU/ g) of recombinant CNTF .

Protein content: Protein quantitation was carried out by two independent methods: 1. UV spectroscopy at 280 nm using the absorbency value of 1.28 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 standard solution of human CNTF as a Reference Standard.

Usage: BioWORLD\'s products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.