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Neurotrophin-3 (NT-3), also known as nerve growth factor-2 (NGF2) or hippocampus-derived neurotro-phic factor (HDNF), is a member of the NGF family of neurotrophic factors and is structurally related to β-NGF, BDNF and NT-4. The NT3 cDNA encodes a 257 amino acid residue precursor protein with a signal peptide and a proprotein that are cleaved to yield the 119 amino acid residue mature NT3.The amino acid sequences of mature human, murine and rat NT-3 are identical. NT-3 selectively promotes the differentiation and survival of specific neuronal subpopulations in both the central as well as the peripheral nervous systems.
Recombinant human NT3 produced in E. coli is a a single, non-glycosylated polypeptide chain containing 119 amino acids and having a molecular mass of 13.6kDa.
Biological activity: The ED50 as determined by the dose-dependant induction of choline acetyl transferase in rat basal forebrain primary septal culture was found between 20-50 ng/ml. The ED50 of NT-3 is typically 0.1-0.2 ng/ml as measured in a cell proliferation assay using a TrkB trans-fected PC12 cell line.
Purity: Greater than 98.0% as determined by:
(a) Analysis by RP-HPLC.
(b) 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 Tyr-Ala-Glu-His-Lys .
Endotoxin: Less than 0.1ng/μg (1.0EU/μg) of rHuNT3 as determined by LAL test.
Sterile and Lyophilized aliquots with a 0.2um filtered protein solution containing 20mM PBS pH7.4.
It is recommended to reconstitute the lyophilized rHuNT3 in sterile PBS not less than 100μg/ml, which can then be further diluted to other solutions.
Lyophilized rHuNT3 although stable at room temperature for 3 weeks, should be stored desiccated below -18°C, which is stable for up to twelve months from date of receipt. Upon reconstitution rHuNT3 should be stored at 4°C between 2-7 days and aliquots of reconstituted samples with 0.1% HSA or BSA are stable up to 3 months below -18°C for future use.
Please avoid freeze-thaw cycles.