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ReliaTech GmbH Specification/DATA SHEET Recombinant Rat Ciliary Neurotrophic Factor (CNTF) 20140814BB FOR RESEARCH ONLY! NOT FOR HUMAN USE! Cat.-no: Size: Lot. No.: Country of origin: R20-001 25 µg According to product label Germany Scientific Background Gene: Cntf Synonyms: Ciliary Neurotrophic Factor, CNTF Ciliary neurotrophic factor (CNTF) is a polypeptide initially purified from chick embryo ocular tissue and identified as a trophic factor for embryonic chick ciliary parasympathetic neurons in culture. Subsequent studies have demonstrated that CNTF is a survival factor for additional neuronal cell types including: dorsal root ganglion sensory neurons, sympathetic ganglion neurons, embryonic motor neurons, major pelvic ganglion neurons and hippocampal neurons. CNTF has also been shown to prevent the degeneration of motor axons after axotomy. The cDNA for CNTF encodes a 199 amino acid residue polypeptide that lacks a signal sequence. CNTF is highly conserved across species and exhibits cross-species activities. Human and rat CNTF share approximately 83% homology in their protein sequence. CNTF is structurally related to IL6, IL11, LIF, and OSM. All of these four helix bundle cytokines share gp130 as a signal transducing subunit in their receptor complexes. . Sequence AFAEQTPLTLHRRDLCSRSIWLARKIRSDLTALMESYVKHQGLNKNINLDSV DGVPVASTDRWSEMTEAERLQENLQAYRTFQGMLTKLLEDQRVHFTPTEGDF HQAIHTLMLQVSAFAYQLEELMVLLEQKIPENEADGMPATVGDGGLFEKKLW GLKVLQELSQWTVRSIHDLRVISSHQMGISALESHYGAKDKQM Database References Protein RefSeq: NP_037298.1 Uniprot ID: P20294 mRNA RefSeq: NM_013166.1 Product Specifications Expressed in E.coli Purity > 98% by SDS-PAGE & silver stain Endotoxin < 0.1ng per µg of CNTF Buffer 5 mM sodium acetate, pH 6.5 Stabilizer None Formulation lyophilized Length (aa): 199 MW: 22,7 kDa Result by Nterminal sequencing AFAEQTPLTHR Stability: The lyophilized powder although stable at room temperature for 3 weeks, is best stored desiccated at -20°C. Reconstituted rat CNTF should be stored in working aliquots at – 20°C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA). Reconstitution: Rat CNTF should be reconstituted in PBS to a concentration of 0.1 mg/ml. This solution can be diluted in water or other buffer solutions or stored at -20°C. AVOID REPEATED FREEZE AND THAW CYCLES! References 1. Finn T and Nishi R, Perspect Dev Neurobiol 4, 1996. 2. Sendtner M et al, J Neurol Sci 124 Suppl:77-83, 1994. 3. Sendtner M et al, J Cell Sci Suppl 15, 1991. Biological Activity: Measured in a cell proliferation assay using TF1 human erythroleukemic cells [Kitamura T et al, J Cell Physiol 140, 1989). The ED50 for this effect is typically 3 - 15 ng/mL. ReliaTech GmbH Phone: +49 (0)5331 8586 987 Fax: +49 (0)5331 8586 989 E-mail: [email protected] web: www.reliatech.de Location: Lindener Str. 15, 38300 Wolfenbüttel, Germany 1 ReliaTech GmbH Specification/DATA SHEET Recombinant Rat Ciliary Neurotrophic Factor (CNTF) rC NT F Handling/Application M 50 40 - 30 - 25 20 - 15 (SDS-PAGE 15%, red.) Fig. 1: SDS-PAGE analysis of recombinant rat CNTF. Sample was loaded in 15% SDS-polyacrylamide gel under reducing condition and stained with Coomassie blue. cells/ml (% of control) 250 200 150 100 RELIATech Competitor 0 5 10 15 20 25 rCNTF [ng/ml] Fig. 2: Proliferation assay with TF-1 cells. ReliaTech GmbH Phone: +49 (0)5331 8586 987 Fax: +49 (0)5331 8586 989 E-mail: [email protected] web: www.reliatech.de Location: Lindener Str. 15, 38300 Wolfenbüttel, Germany 2