Mouse beta Nerve Growth Factor Recombinant

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Mouse beta Nerve Growth Factor Recombinant

$70.00$2,700.00


accession P01139


Source Optimized DNA sequence encodingMouseNerve Growth Factor mature chain was expressed in CHO.
Molecular weight NativeMouse Nerve Growth Factor is generated by the proteolytic removal of the signal peptide and propeptide the molecule has a calculated molecular mass of approximately kDa. RecombinantMouse Nerve Growth Factor is a monomeric protein consisting of amino acid residue subunits, and migrates as an approximately13 kDa protein under non-reducing and reducing conditions in SDS-PAGE.
Purity >95%, as determined by SDS-PAGE and HPLC
Biological Activity The ED(50) was determined by the determined by its ability to stimulate chick E9 DRG neurite outgrowth was found to be in the range of1.0 ng/ml.

Protein Sequence MSMLFYTLIT AFLIGVQAEP YTDSNVPEGD SVPEAHWTKL QHSLDTALRR ARSAPTAPIA ARVTGQTRNI TVDPRLFKKR RLHSPRVLFS TQPPPTSSDT LDLDFQAHGT IPFNRTHRSK RSSTHPVFHM GEFSVCDSVS VWVGDKTTAT DIKGKEVTVL AEVNINNSVF RQYFFETKCR ASNPVESGCR GIDSKHWNSY CTTTHTFVKA LTTDEKQAAW RFIRIDTACV CVLSRKATRR G
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 Recombinant mouse NGF was lyophilized from a.2 μm filtered solution inmM sodium acetate, 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 P07174
Molecular function Growth-factor
Molecular function Metalloenzyme-inhibitor
Molecular function Protease-inhibitor

Methods

Cell culture

  • DRGs from female Wistar rats (150 grams) were harvested in cold DMEM and any excess dorsal roots and spinal nerves were trimmed under a stereo microscope.
  • DRGs were incubated with 0.125% Collagenase XI and 0.1 mg/ml DNase II in DMEM for 90 min at 37°C.
  • After enzymatic digestion, DRGs were triturated with a cut 1 ml tip until a cell suspension was obtained.
  • Cells were spun down, resuspended in pre-warmed DMEM and filtered through a 70 µm mesh .
  • DRG neurons were recovered by using 10% BSA (PAA)/ DMEM cushions and plated on 13 mm glass coverslips coated with poly-L-lysine and laminin in complete media (DMEM, 10% fetal bovine serum (FBS), penicillin/streptomycin (1∶100), NGF (50 ng/ml) and aphidicolin (10 µM).
  • 10,000 and 150,000 cells per coverslips were plated for immunofluorescence and biochemistry purposes, respectively.
  • Cells were maintained in a 95% air/5% CO2 humidified incubator.

NGF contents in the skin.

NGF
  • NGF contents in the skin of the affected and contralateral hindlimbs of wild-type and Y1472F-KI mice on day 50 post-inoculation and in the back skin at 24 h after wounding were measured by ELISA.