Human Bone Morphogenetic protein-2 Recombinant
Categories: TGF-beta familyTGF-beta familyRecombinant Human Cytokines$70.00 – $4,700.00
Description
Accession
P12643
Source
Optimized DNA sequence encoding Human Bone Morphogenetic Protein-2 mature chain was expressed in Escherichia Coli.
Molecular weight
Native human Bone Morphogenetic Protein-2 is generated by the proteolytic removal of the signal peptide and propeptide, the molecule has a calculated molecular mass of approximately 13 kDa. Recombinant BMP-2 is a disulfide-linked homodimeric protein consisting of two 115 amino acid residue subunits.BMP2 migrates as an approximately 25 kDa protein under non-reducing conditions and as a 13 kDa protein under reducing conditions in SDS-PAGE.
Purity
>95%, as determined by SDS-PAGE and HPLC
Biological Activity
The ED(50) was determinedby its ability to induce alkaline phosphatase production by C2C12 myogenic cells cells was found to be less than ng/ml.
Protein Sequence
MVAGTRCLLA LLLPQVLLGG AAGLVPELGR RKFAAASSGR PSSQPSDEVL SEFELRLLSM FGLKQRPTPS RDAVVPPYML DLYRRHSGQP GSPAPDHRLE RAASRANTVR SFHHEESLEE LPETSGKTTR RFFFNLSSIP TEEFITSAEL QVFREQMQDA LGNNSSFHHR INIYEIIKPA TANSKFPVTR LLDTRLVNQN ASRWESFDVT PAVMRWTAQG HANHGFVVEV AHLEEKQGVS KRHVRISRSL HQDEHSWSQI RPLLVTFGHD GKGHPLHKRE KRQAKHKQRK RLKSSCKRHP LYVDFSDVGW NDWIVAPPGY HAFYCHGECP FPLADHLNST NHAIVQTLVN SVNSKIPKAC CVPTELSAIS MLYLDENEKV VLKNYQDMVV EGCGCR
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 Human Bone Morphogenetic Protein-2 was lyophilized from a 0.2 μm filtered solution inmM AcOH 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
O88273
Interactor
Interactor
Interactor
Interactor
Interactor
Interactor
P97798
Biological Process
Biological Process
Biological Process
Molecular function
Molecular function
Molecular function
Methods
BMP-2 induces cell migration by increasing dimensions of nanowaves.
- (a) Immunofluorescence staining of migrating pre-osteoblastic cells without and with BMP-2 after culturing for 12 h; the cells were stained for visualization of the actin filaments (green).
In vitro osteogenic differentiation
- Primary osteoblast precursor cells were isolated from the calvarial bone from newborn mice by enzymatic digestion with α-MEM containing collagenase and 0.2% dispase II .
- The precursor cells were plated at a density of 2 × 104 cells/well in 48-well plates and cultured for 7–14 days in medium'>osteogenic medium containing 10 mM β-glycerol phosphate, 50 μg/ml ascorbate-2-phosphate, 10−7μM dexamethasone and 25 ng/ml human recombinant BMP-2 .
- The culture medium was replaced with fresh osteogenic medium every 3 days.
- Alkaline phosphatase (ALP) staining was performed by BCIP/NBT color development substrate according to the manufacturer’s instructions.
- ALP activity assay was measured biochemically using a
p nitrophenyl phosphate tablets . - Cells were differentiated for 7 days and harvested.
- The cells were then lysed with 1% Triton X-100.
- Lysates were centrifuged at 13,000 ×
g for 10 min at 4°C. - Cell extracts were added to each 96-well plate containing
p -nitrophenyl phosphate and incubated for 30 min at…
Determination of Cell Response
- Conditioned media were collected and levels of secreted osteocalcin , osteoprotegerin , bone morphogenetic protein-2 (BMP2) (, ocky , ), BMP4 , vascular endothelial growth factor A (VEGFA) , and fibroblast growth factor-2 (FGF2) were measured by immunoassay following manufacturer’s instructions.
