Active Bone Morphogenetic Protein 2 (BMP2)

BMP2A; BMP-2A; Hemochromatosis Modifier

ACTIVITY TEST

Bone morphogenetic protein 2(BMP2) belongs to the TGF-β superfamily of proteins. It plays an important role in the development of bone and cartilage. It is involved in the hedgehog pathway, TGF beta signaling pathway, and in cytokine-cytokine receptor interaction. BMP2 is also involved in cardiac cell differentiation and epithelial to mesenchymal transition. Like many other proteins from the BMP family, BMP2 has been demonstrated to potently induce osteoblast differentiation in a variety of cell types. Besides, Noggin (NOG) has been identified as an interactor of BMP2, thus a binding ELISA assay was conducted to detect the interaction of recombinant human BMP2 and recombinant human NOG. Briefly, BMP2 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100ul were then transferred to NOG-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-BMP2 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution , wells were incubated 15-25 minutes at 37℃. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of BMP2 and NOG was shown in Figure 1, and this effect was in a dose dependent manner.

To test the effect of BMP2 on cell apoptosis, HepG2 cells were seeded into triplicate wells of 96-well plates at a density of 4,000 cells/well and allowed to attach overnight, then the medium was replaced with various concentrations of recombinant human BMP2 diluted with 5% serum standard DMEM. After incubated for 72h, cells were observed by inverted microscope and cell proliferation was measured by Cell Counting Kit-8 (CCK-8). Briefly, 10 µl of CCK-8 solution was added to each well of the plate, then the absorbance at 450 nm was measured using a microplate reader after incubating the plate for 1-4 hours at 37 ℃. Apoptosis of HepG2 cells after incubation with BMP2 for 72h observed by inverted microscope was shown in Figure 1. Cell viability was assessed by CCK-8 (Cell Counting Kit-8 ) assay after incubation with recombinant human BMP2 for 72h. The result was shown in Figure 2. It was obvious that BMP2 significantly decreased cell viability of HepG2 cells. The ED50 of recombinant human BMP2 is 3.25 ug/ml.

USAGE

Reconstitute in 20mM Tris, 150mM NaCl (PH8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
Journal of Applied Toxicology Chronic exposure to low concentrations of strontium 90 affects bone physiology but not the hematopoietic system in mice Wiley: Source
PLoS ONE A Combinatorial Relative Mass Value Evaluation of Endogenous Bioactive Proteins in Three-Dimensional Cultured Nucleus Pulposus Cells of Herniated Intervertebral Discs: Identification of Potential Target Proteins for Gene Therapeutic Approaches Plosone: Source
Medical Journal of Chinese People's Liberation Army Effect of transplantation of BMP-2-induced bone marrow mesenchymal stem cells on myocardial infarction of rats after reperfusion Source
PLOS ONE PDGF-AA promotes osteogenic differentiation and migration of mesenchymal stem cell by down-regulating PDGFRα and derepressing BMP-Smad1/5/8 signaling Pubmed:25470749
Eur J Pharmacol Negative effect of serotonin–norepinephrine reuptake inhibitor therapy on rat bone tissue after orchidectomy PubMed: 25934570
Pharmacology Effect of Mirtazapine on Rat Bone Tissue after Orchidectomy PubMed: 25871861
Journal of Cellular Biochemistry Titanium with Nanotopography Induces Osteoblast Differentiation by Regulating Endogenous Bone Morphogenetic Protein Expression and Signaling Pathway PubMed: 26681207
International Journal of Nanomedicine Sustained dual release of placental growth factor-2 and bone morphogenic protein-2 from heparin-based nanocomplexes for direct osteogenesis Pubmed:27042064
Research report EFFECTS OF FLUORIDE ON THE EXPRESSION OF BMP-2 AND SMAD1 IN RAT OSTEOBLASTS IN VITRO files:p013-022
Gene  Unveiling the interactions among BMPR-2, ALK-1 and 5-HTT genes in the pathophysiology of HAPE Pubmed:27196063
Journal of Materials Chemistry B Extracellular Matrix Powder from Cultured Cartilage-Like Tissue as Cell Carriers for Cartilage Repair DOI:10.1039/C7TB00640C
Scientific Reports 3D- Printed Poly(ε-caprolactone) Scaffold Integrated with Cell-laden Chitosan Hydrogels for Bone Tissue Engineering. pubmed:29042614
Journal of Applied Biomaterials & Functional Materials Demineralized dentin and enamel matrices as suitable substrates for bone regeneration pubmed:28731486
Journal of Clinical Periodontology  Peroxisome proliferator‐activated receptor γ plays dual roles on experimental periodontitis in rats Pubmed:29574908
Pharmacology Effects of Amlodipine on Bone Metabolism in Orchidectomised Spontaneously Hypertensive Rats Pubmed:29898457
Journal of Applied Biomaterials & Functional Materials BMP-2 and type I collagen preservation in human deciduous teeth after demineralization Pubmed:30045659
Applied Surface Science Enhanced osteogenic differentiation of MC3T3-E1 on rhBMP-2 immobilized titanium surface through polymer-mediated electrostatic interaction
Materials Science and Engineering: C Injectable hydrogels from enzyme-catalyzed crosslinking as BMSCs-laden scaffold for bone repair and regeneration
Biomaterials Science Development of CaCO 3 microsphere-based composite hydrogel for dual delivery of growth factor and Ca to enhance bone regeneration
ENVIRONMENTAL TOXICOLOGY AND PHARMACOLOGY The dose-time effects of fluoride on the expression and DNA methylation level of the promoter region of BMP-2 and BMP-7 in rats Pubmed: 32004919
Journal of Industrial and Engineering Chemistry Biofabrication and application of decellularized bone extracellular matrix for effective bone regeneration
BMC Musculoskeletal Disorders Synergistic Effect of Platelet-rich Fibrin Releasate and Bone Marrow Stem Cells in Preventing Bone Loss in Ovariectomized Mouse
Bioact Mater New design to remove leukocytes from platelet-rich plasma (PRP) based on cell dimension rather than density 33842739
Tissue Eng Regen Med Osteogenic and Angiogenic Potency of VEGF165-Transfected Canine Bone Marrow Mesenchymal Cells Combined with Coral Hydroxyapatite in Vitro 34302695
Toxicon The WNT/β-catenin signalling pathway induces chondrocyte apoptosis in the cartilage injury caused by T-2 toxin in rats 34742779
ACS Biomater Sci Eng Polydopamine-Coated Poly(l-lactide) Nanofibers with Controlled Release of VEGF and BMP-2 as a Regenerative Periosteum 34472855
Phytomedicine Efficacy of Total flavonoids of Rhizoma drynariae on the Blood Vessels and the Growth Quality of Bone Graft in the Induced Membrane Pubmed:35278899
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