Mouse beta Nerve Growth Factor Recombinant
Categories: Neurotrophic factorsRecombinant Mouse Cytokines$70.00 – $2,700.00
Description
Accession
P01139
Source
Optimized DNA sequence encoding Mouse Nerve Growth Factor mature chain was expressed in CHO.
Molecular weight
Native Mouse beta Nerve Growth Factor is generated by the proteolytic removal of the signal peptide and propeptide the molecule has a calculated molecular mass of approximately 13 kDa. Recombinant Mouse beta Nerve Growth Factor is a homodimeric protein consisting of 2x120 amino acid residue subunits, and migrates as an approximately 26 kDa protein under non-reducing and 13kDa 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 0.1 ng/µg(1EU/µg).
Presentation
Recombinant mouse NGF was lyophilized from a 0.2 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 0.1 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 at2° -8° 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
Molecular function
Molecular function
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 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.
