Human Brain-Derived Neurotrophic Factor Recombinant
Categories: Neurotrophic factorsRecombinant Human Cytokines$70.00 – $4,700.00
Description
Accession
P23560
Source
Optimized DNA sequence encoding Human Brain Derived Neurotrophic Factor mature chain was expressed in Escherichia Coli.
Molecular weight
Human native BDNF is generated by the proteolytic removal of the signal peptide and propeptide, the molecule has a calculated molecular mass of approximately 14 kDa. Recombinant human BDNF is a disulfide-linked homodimeric protein consisting of two 119 amino acid residue subunits. migrates as an approximately 27 kDa protein under non-reducing conditions and as a 14 kDa protein under reducing conditions in SDS-PAGE.
Purity
>97%, as determined by SDS-PAGE and HPLC
Biological Activity
The ED(50) was determined by the dose-dependent proliferation of rat C6 cells, and was found to be in the range of 0.5 ug/ml.
Protein Sequence
MTILFLTMVI SYFGCMKAAP MKEANIRGQG GLAYPGVRTH GTLESVNGPK AGSRGLTSLA DTFEHVIEEL LDEDQKVRPN EENNKDADLY TSRVMLSSQV PLEPPLLFLL EEYKNYLDAA NMSMRVRRHS DPARRGELSV CDSISEWVTA ADKKTAVDMS GGTVTVLEKV PVSKGQLKQY FYETKCNPMG YTKEGCRGID KRHWNSQCRT TQSYVRALTM DSKKRIGWRF IRIDTSCVCT LTIKRGR
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 Brain Derived Neurotrophic Factorwas lyophilized from a 0.2 μm filtered citric acid, pH.7.
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
Interactor
Interactor
Interactor
Q99523
Molecular function
Methods
Effects of BDNF on RANKL expression in human bone marrow stromal cells (BMSCs).
- The dose and time responses to BDNF of RANKL mRNA expression in BMSCs.
Expression of key growth factors in human endometrium.
- Brown signals for growth factors (EGF, IGF-1, GM-CSF, BDNF, CSF-1, artemin, and GDNF) were found following staining with specific antibodies.
Expression of key growth factors in human endometrium.
- Brown signals for growth factors (EGF, IGF-1, GM-CSF, BDNF, CSF-1, artemin, and GDNF) were found following staining with specific antibodies.
Surgery and Drug Microinjection
- For drug microinjection, the stylus was removed from the guide cannula, and a 28-gauge microinjection cannula (extending 1.0 mm from the tip) was inserted, which was attached via polyethylene tubing to a Hamilton microsyringe driven by an injection pump (KD Science, KDS310, US).
- K252a (, , 25 µM, 1 µl/side)
TCT TCC CCT TTT AAT GGT 3′; BDNF missense oligonucleotide (MSO): 5′ATA CTT TCT GTT CTT GCC 3′, 2 nM, 0.5 µl/side] were dissolved in sterile saline
BDNF tetrapeptides used in this study.
- Amino acid sequence of human BDNF.
Ischemia induces BDNF mRNA expression by neuronal and endothelial cells in the ipsilateral striatum.
- PCR analysis of BDNF expression in the ischemic striatum 1 week after MCAo.
Differentiation of smNPCs
- For generation of more ventral CNS neurons, including mDANs, medium'>smNPC expansion medium was changed 2 days after splitting to N2B27 medium with 100 ng/mL FGF8 , 1 µM PMA, and 200 µM AA.
- After 8 days in this medium, maturation medium–N2B27 with 10 ng/mL BDNF , 10 ng/mL GDNF , 1 ng/mL TGF-b3 , 200 µM AA, and 500 µM dbcAMP–was used for the maturation of neurons.
- 0.5 µM PMA was added to this medium for 2 more days.
- One day after changing to maturation medium, the cultures were split at a 1∶3 ratio as small clumps, or single cells after Accutase treatment, or earlier when cultures became over-confluent.
- Cultures were analyzed after 2 weeks in maturation conditions unless otherwise indicated.
Differentiation of smNPCs
- For generation of more ventral CNS neurons, including mDANs, medium'>smNPC expansion medium was changed 2 days after splitting to N2B27 medium with 100 ng/mL FGF8 , 1 µM PMA, and 200 µM AA.
- After 8 days in this medium, maturation medium–N2B27 with 10 ng/mL BDNF , 10 ng/mL GDNF , 1 ng/mL TGF-b3 , 200 µM AA, and 500 µM dbcAMP–was used for the maturation of neurons.
- 0.5 µM PMA was added to this medium for 2 more days.
- One day after changing to maturation medium, the cultures were split at a 1∶3 ratio as small clumps, or single cells after Accutase treatment, or earlier when cultures became over-confluent.
- Cultures were analyzed after 2 weeks in maturation conditions unless otherwise indicated.
Primary culture of mouse retinal ganglion cells
-
C57BL/6 J or Dock3-Tg mice (7–10 days old) were used for primary culture of RGCs according to Winzeler
et al. [g for 10 min. - RGCs were suspended in medium'>medium containing 1 mM glutamine, 5 μg/ml insulin, 60 μg/ml N-acetylcysteine, 62 ng/ml progesterone, 16 μg/ml putrescine, 40 ng/ml sodium selenite, 0.1 mg/ml BSA, 40 ng/ml triiodothyronine, 0.1 mg/ml transferrin, 1 mM sodium pyruvate, 2% B27 supplement , 10 μM forskolin , 50 ng/ml brain-derived neurotrophic factor (BDNF, , ), 50 ng/ml ciliary neurotrophic factor (CNTF), and 50 ng/ml basic fibroblast growth factor (bFGF) in medium'>Neurobasal medium'>medium .
- Ninety-six–well culture plates were coated with poly-D-lysine and laminin and mouse RGCs were plated at a density of 4,000 cells/well (or 4,000 cells/culture insert for μ-dishes (ibidi)) and cultured for at least 10 days before the experiments.
Induced Differentiation of Muse-ATs
- For medium'>medium'>neural cell formation, Muse cells-AT were incubated as non-adherent cells in ultra-low attachment plates (Corning Incorporated , , ) in the presence of medium'>medium'>neural medium'>medium'>differentiation medium'>medium 1 containing medium'>Neurobasal medium'>medium supplemented with B-27 supplement serum free , 100 µg/ml kanamycin , 2 mM glutamine , 30 ng/ml bFGF and 30 ng/ml EGF for 7 days
