Artemin, Human Recombinant Human Artemin
Total Page:16
File Type:pdf, Size:1020Kb
Artemin, human Recombinant Human Artemin Instruction Manual Catalog Number C-60031 Synonyms ART, ARTN , EVN, NBN Description Artemin is a disulfide-linked homodimeric neurotrophic factor structurally related to GDNF, Artemin, Neurturin and Persephin. These proteins belong to the cysteine-knot superfamily of growth factors that assume stable dimeric protein structures. Artemin, GDNF, Persephin and Neurturin all signal though a multicomponent receptor system, composed of RET (receptor tyrosine kinase) and one of the four GFR-alpha (alpha1-alpha4) receptors. Artemin prefers the receptor GFRalpha3-RET, but will use other receptors as an alternative. Artemin supports the survival of all peripheral ganglia such as sympathetic, neural crest and placodally derived sensory neurons, and dompaminergic midbrains neurons. The functional human Artemin ligand is a disulfide-linked homodimer, of two 12.0 kDa polypeptide monomers. Each monomer contains seven conserved cysteine residues, one of which is used for inter-chain disulfide bridging and the others are involved in intramolecular ring formation known as the cysteine knot configuration. Recombinant human Artemin is a 24.2 kDa, disulfide-linked non-glycosylated homodimer formed by two identical 113 amino acid subunits. Recombinant Artemin has been purified using proprietary chromatographic techniques. Quantity 20 µg Molecular Mass 24.2 kDa Source E. coli Biological-Activity Artemin is fully biologically active when compared to a standard. Assay #1: Determined by its ability to stimulate the proliferation of human SH-SY5Y cells. The expected EDЊЅ for this effect is 2.0-5.0 ng/ml. Assay #2: Determined by its ability to promote neuronal survival and neurite outgrowth on dorsal root ganglion neurons. Specific Activity see below Formulation Sterile-filtered through a 0.2 micron filter and lyophilized from 10 mM sodium citrate buffer (pH 4.5), 25 mM NaCl. Reconstitution Please Note: Always centrifuge product briefly before opening vial. The lyophilized protein should be reconstituted in sterile, ultra-pure water to a concentration of 0.1-1.0 mg/ml. This solution can then be diluted into other aqueous buffers and stored at -20°C for future use. Purity Greater than 98% (determined by SDS-PAGE and RP-HPLC analysis). Endotoxin Level < 0.1 ng per µg of rHu Artemin (1EU/µg). Storage The lyophilized protein, though stable at room temperature for up to 3 weeks, is best stored desiccated at -20°C. Reconstituted rHu Artemin should be used immediately or stored long-term in undiluted working aliquots at -20°C. For long-term storage it is recommended to add a carrier protein (0.1% endotoxin-free HSA or BSA; e.g. Cat.No. C-69500A). Avoid repeated freeze-thaw cycles. 05/2019.