Ciliary Neurotrophic Factor (human)
PRODUCT INFORMATION
Cat. No.: Z00332
Size: 1 mg or 20 ug or 5 ug
Source: Escherichia Coli.
Purity：
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 22,706 Dalton. The
rHuCNTF is purified by proprietary chromatographic techniques. Ciliary neurotrophic
factor was first identified as a survival factor for neurons from the ciliary ganglion of
chicken embryos. Most of its known actions are restricted to cells of the nervous
system, including motor neurons, sympathetic ganglion neurons, sensory neurons,
hippocampal neurons, and medial septal neurons. CNTF also prevents degeneration
of motor axons after axotomy and promotes astrocyte differentiation and
oligodendrocyte survival and maturation. Outside the nervous system, CNTF
maintains embryonic stem cells in an undifferentiated, pluripotent state. CNTF is
structurally related to leukemia inhibitory factor (LIF), interleukin-6 (IL-6), interleukin-11
(IL-11) and oncostatin M (OSM). CNTF exerts its actions through the activation of the
high-affinity CTNF receptor complex, which contains the ligand-binding α-subunit
(CNTF Rα) and two signal transducing β-subunits (LIF Rβ and gp130). CNTF is
localized in the cell nucleus subsequent to receptor binding. Human and rat CNTF
share ~83% sequence homology and show cross-reactivity in bioactivity.
Quantitation
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% (1 mg/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 CNTF as a Reference Standard.
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.
Endotoxin Level:
Less than 0.1 ng/μg (IEU/μg)
of rHuCNTF.
Dimers and aggregates:
Less than 1% as determined
by silver-stained SDS-PAGE
gel analysis.
Sequence analysis:
The sequence of the first five
N-terminal amino acids was
determined and was found to
be Met-Ala-Phe-Thr-Glu.
MW: 22,706 Dalton
Specific Activity
The ED50 as determined by the dose-dependant stimulation of TF-1 cells is < 2 ng/ml,
corresponding to a Specific Activity of 5 x 105 IU/mg.
Storage
Lyophilized rHuCNTF although stable at room temperature for 3 weeks, should be
stored desiccated below -18 °C. Upon reconstitution rHuCNTF 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 prevent
freeze-thaw cycles.
Formulation
Lyophilized from a concentrated (1 mg/ml) solution in water containing 5 mM sodium
Phosphate buffer pH = 7.5 and 5 mM sodium chloride.
Reconstitution
It is recommended to reconstitute the lyophilized rHuCNTF in sterile 18 MΩ-cm H2O not
less than 100 μg/ml, which can then be further diluted to other aqueous solutions.
Sequence
MAFTEHSPLT PHRRDLCSRS IWLARKLRSD LTALTESYVK HQGLNKNINL
DSADGMPVAS TDQWSELTEA ERLQENLQAY RTFHVLLARL LEDQQVHFTP
TEGDFHQAIH TLLLQVAAFA YQIEELMILL EYKIPRNEAD GMPINVGDGG
LFEKKLWGLK VLQELSQWTV RSIHDLRFIS SHQTGIPARG SHYIANNKKM
** For non-clinical research use only. **
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