Human Glial-Derived Neurotrophic Factor Recombinant

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Human Glial-Derived Neurotrophic Factor Recombinant

$70.00$4,700.00


accession P39905


Source Optimized DNA sequence encoding Human GDNF mature chain was expressed in Escherichia Coli.
Molecular weight Native Human Glial-Derived Neurotrophic Factor isgenerated by the proteolytic removal of the signal peptide and propeptide, the molecule has a calculated molecular mass of approximately15 kDa. Recombinant GDNF is a disulfide-linked homodimeric protein consisting of two amino acid residue subunits,and migrates as an approximately30 kDa protein under non-reducingconditions and askDa under reducing conditions in SDS-PAGE.
Purity >95%, as determined by SDS-PAGE and HPLC
Biological Activity The ED(50) was determined by theproliferation of rat C6 cells is <.1 ng/ml, corresponding to a specific activity of > x units/mg.
Protein Sequence MKLWDVVAVC LVLLHTASAF PLPAGKRPPE APAEDRSLGR RRAPFALSSD SNMPEDYPDQ FDDVMDFIQA TIKRLKRSPD KQMAVLPRRE RNRQAAAANP ENSRGKGRRG QRGKNRGCVL TAIHLNVTDL GLGYETKEEL IFRYCSGSCD AAETTYDKIL KNLSRNRRLV SDKVGQACCR PIAFDDDLSF LDDNLVYHIL RKHSAKRCGC I
Endotoxin Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than.1 ng/µg(1EU/µg).
Presentation RecombinantGlial-Derived Neurotrophic Factor was lyophilized from a.2 μm filtered solution in.5% glycine,.5% sucrose,.01% Tween80, mM Glutamic acid, pH.5.
Reconstitution A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than.1 mg/mL. This solution can then be diluted into other buffers.
Storage The lyophilized protein is stable for at least years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at° -° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
Usage This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.


Interactor Q8N5Y2
Interactor P56159 GFRA1_HUMAN
Molecular function Growth-factor

Methods

Overexpression of EpICD in ROSA GS cells.

GDNF
  • Upregulation of EPCAM by GDNF stimulation.

Surface Plasmon resonance (Biacore) measurements

  • MCer1 was immobilized on a CM5 chip in BIAcore 2000 with an amine group.
  • Binding of Cer1 to hBMP2 , hBMP4, or hGNF (1 µg/ml& ) was analysed in triplicate at 25°C in HBS-P buffer supplemented with 10 mM HEPES, 150 mM NaCl and 0.005% Tween 20 at pH 7.4, with a flow rate of 20 µg/ml min.
  • The kinetics and the dissociation constant (KD) were calculated with BIAevaluation software ver.
  • 4.1 .

Preparation of Primary Hippocampal Cultures

  • Primary hippocampal cultures were prepared from wild-type neonatal (E19) rat embryos (timed pregnant Sprague Dawley rats were obtained from Charles River Laboratories, Wilmington, MA) as described previously

RET51 internalizes more efficiently than RET9.

GDNF
  • Retinoic acid–treated SH-SY5Y cells were serum starved, incubated with or without 100 ng/ml GDNF for 20 min, and lysed.

RET51 internalizes more efficiently than RET9.

100 ng/ml GDNF
  • Retinoic acid–treated SH-SY5Y cells were serum starved, incubated with or without 100 ng/ml GDNF for 20 min, and lysed.