BMP-7 (osteogenic protein 1) belongs to the Bone-growth regulatory factors that are members of the transforming growth factor-beta (TGF-beta) superfamily of proteins. They are synthesized as large precursor molecules which are cleaved by proteolytic enzymes. The active form can consist of a dimer of two identical proteins or a heterodimer of two related bone morphogenetic proteins. N-TERMINAL---Human BMP-2 (Met 1 Arg 282) Human BMP-7 (Ser 293 Arg 431)---C-TERMINAL. The DNA sequence encoding the human BMP-2 signal peptide and propeptide (1~282 amino acid) fused to the human BMP-7 mature chain (293~431 amino acid) was expressed in a Chinese hamster ovary cell line. The mature recombinant BMP-7 generated by the proteolytic removal of the signal peptide and propetide contains 139 amino acid residues. The glycosylation of BMP-7 increases the molecular mass and the glycosylated proteins migrate as 25 ~ 40 kDa in SDS-PAGE under non-reducing conditions.
Protein Details
Purity
>95% by SDS Page and analyzed by silver stain.
Endotoxin Level
<1.0 EU/µg as determined by the LAL method
Biological Activity
The biological activity of Human BMP-7 was determined by its ability to induce alkaline phosphatase production by mouse ATDC5 chondrogenic cells (Nakamura, K. et al., 1999, Exp. Cell Res. 250:351). The expected ED<sub>50</sub> is typically 0.2 - 0.6 μg/mL.
Amino Acid Sequence
The DNA sequence encoding the human BMP-2 signal peptide and propeptide (amino acid residues 1 - 282 of human BMP-2) fused to the human BMP-7 mature chain (amino acid residues 293 - 431 of human BMP-7) cloned into a suitable mammalian expression vector and expressed in a Chinese hamster ovary cell line.
State of Matter
Lyophilized
Predicted Molecular Mass
The molecular weight of Recombinant Human BMP-7, due to glycosylation is approximately 30 - 40 kDa under non-reducing conditions and runs as an 18 - 20 kDa protein under reducing conditions.
Formulation
This recombinant protein was lyophilized from a 0.2 μm filtered solution in 33% acetonitrile (CH3CN) and 0.1% trifluoroacetic acid (TFA).
Storage and Stability
The lyophilized protein should be stored desiccated at -20°C. The reconstituted protein can be stored for at least one week at 4°C. For long-term storage of the reconstituted protein, aliquot into working volumes and store at -20°C in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles.
Country of Origin
USA
Shipping
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Recombinant Human BMP-7 (CHO Cell Expressed) is widely used in research due to its well-characterized biological activity, post-translational modifications, and relevance in multiple physiological and pathological processes.
Key scientific reasons to use this protein in your research applications:
Functional Activity: BMP-7 is a member of the TGF-β superfamily and is crucial for bone formation, cartilage induction, nephron development, and neuroregeneration. It also plays roles in inhibiting epithelial-mesenchymal transition (EMT), reducing fibrosis, and modulating inflammation.
CHO Cell Expression: Proteins expressed in CHO (Chinese Hamster Ovary) cells undergo mammalian glycosylation and folding, closely mimicking native human BMP-7. This ensures biological activity and stability suitable for in vitro and in vivo studies. CHO-expressed BMP-7 is less likely to be immunogenic compared to proteins expressed in bacterial systems, making it preferable for translational and therapeutic research.
Applications: Recombinant BMP-7 is used in:
Bone and cartilage tissue engineering: Induces osteogenic and chondrogenic differentiation.
Fibrosis and inflammation studies: Antagonizes TGF-β signaling, reduces fibrotic markers, and promotes anti-inflammatory macrophage polarization.
Cardiovascular research: Attenuates pathological cardiac remodeling and hypertrophy.
Nephrology: Ameliorates fibrotic damage in kidney disease models.
Neurobiology: Promotes neuroregeneration after injury.
Experimental Consistency: CHO-expressed recombinant BMP-7 offers batch-to-batch consistency and is validated for use in functional assays, bioassays, and cell culture applications.
Clinical Relevance: Recombinant human BMP-7 produced in CHO cells has been used clinically for bone repair and spinal fusion, supporting its translational potential.
Summary of best practices:
Use CHO cell-expressed BMP-7 for experiments requiring native-like glycosylation and folding.
Validate protein activity in your specific assay system.
Employ in studies of bone/cartilage formation, fibrosis, inflammation, and tissue regeneration.
In conclusion, recombinant human BMP-7 (CHO cell expressed) is a scientifically robust choice for research applications requiring high-quality, biologically active protein with mammalian post-translational modifications, especially in fields related to tissue engineering, fibrosis, inflammation, and regenerative medicine.
Yes, you can use Recombinant Human BMP-7 (CHO cell-expressed) as a standard for quantification or calibration in your ELISA assays, provided the ELISA kit is validated for detecting recombinant BMP-7 and the recombinant protein is compatible with the kit's detection antibodies.
Key Points:
R&D Systems Quantikine Human BMP-7 ELISA Kit (DBP700): This kit explicitly states that it contains CHO cell-expressed recombinant human BMP-7 and is designed to accurately quantitate both natural and recombinant human BMP-7. Therefore, using CHO-expressed recombinant BMP-7 as a standard is appropriate and recommended for this kit.
Other ELISA Kits: Some kits may not be validated for recombinant proteins, especially if the recombinant protein is expressed in a different system (e.g., E. coli, yeast) or has different post-translational modifications. Always check the manufacturer’s documentation for compatibility.
General Best Practice: When using recombinant protein as a standard, ensure:
The recombinant protein is expressed in the same system as the standard provided by the kit (e.g., CHO cells).
The protein is purified and characterized for activity and purity.
The standard curve is prepared in the same matrix as your samples to avoid matrix effects.
References:
R&D Systems Quantikine Human BMP-7 ELISA Kit (DBP700) documentation confirms the use of CHO-expressed recombinant human BMP-7 as a standard.
Always consult the specific kit’s manual for validation and compatibility information.
If you are using a different ELISA kit, verify with the manufacturer or technical support to ensure compatibility with your recombinant standard.
Recombinant Human BMP-7 (CHO Cell Expressed) has been validated in published research for a range of applications, primarily focused on its biological activity in cell-based assays, bone and cartilage regeneration, stem cell differentiation, and functional bioassays.
Key validated applications include:
Bioassays: BMP-7 is widely used to assess its ability to induce cellular responses such as alkaline phosphatase production in chondrogenic cells, osteogenic differentiation, and other growth factor activities in various cell types (e.g., mouse ATDC5 chondrogenic cells, human bone marrow stromal cells).
Stem Cell Differentiation: BMP-7 has been used to promote osteogenic and chondrogenic differentiation of mesenchymal stem cells, as well as to study early trophectoderm states and hepatocyte-like cell differentiation from induced pluripotent stem cells.
Bone and Cartilage Regeneration: Validated for in vitro and in vivo studies of bone formation, fracture healing, spinal fusion, and cartilage repair. BMP-7 stimulates maturation of osteoblastic progenitors and induces differentiation of non-osteogenic cells to osteoblasts.
Functional Assays: Used to evaluate growth factor activity, including effects on insulin sensitivity, neuronal regeneration, and endothelial cell function.
ELISA Capture and Surface Plasmon Resonance (SPR): BMP-7 has been used as a capture reagent in immunoassays and for studying protein-protein interactions.
In Vivo Studies: Demonstrated efficacy in animal models for bone repair, diabetes-related insulin sensitivity, and spinal cord injury recovery.
Implant Functionalization: BMP-7 has been used to coat biomaterials (e.g., titanium dental implants) to enhance osseointegration and tissue regeneration.
Additional validated research contexts:
Chondrocyte Redifferentiation: BMP-7, alone or in combination with other growth factors (e.g., TGF-β3), stimulates chondrocyte redifferentiation under various oxygen conditions.
Cancer Research: Used to study stemness and metastasis in cancer models, such as prostate cancer and medulloblastoma tumorigenesis.
Cell Signaling Studies: BMP-7 has been validated for investigating canonical and non-canonical signaling pathways, including P38 MAP kinase and SOX family gene activation.
In summary, CHO cell-expressed recombinant human BMP-7 is validated for:
In vivo animal models for tissue regeneration and metabolic studies
Biomaterial and implant functionalization for enhanced tissue integration.
These applications are supported by multiple peer-reviewed publications and product validation data, confirming the protein’s utility in both basic research and translational studies.
To reconstitute and prepare Recombinant Human BMP-7 (CHO Cell Expressed) for cell culture experiments, follow these best-practice steps:
Centrifuge the vial briefly (e.g., 20–30 seconds in a microcentrifuge) before opening to ensure all lyophilized protein is at the bottom.
Warm the vial to room temperature before opening to avoid condensation.
Reconstitution:
Use a suitable solvent. Common options include:
Sterile 10–20 mM acetic acid (AcOH).
Sterile water containing 0.1% human serum albumin (HSA) or bovine serum albumin (BSA) to stabilize the protein and prevent adsorption to surfaces.
Some protocols recommend sterile 4 mM HCl with 0.1% carrier protein.
Recommended concentration: Reconstitute to at least 100 μg/mL (0.1 mg/mL), with some protocols suggesting up to 0.2 mg/mL or 1 mg/mL depending on your application.
Mix gently: Do not vortex or shake vigorously. Gently swirl or tap the vial to dissolve the protein.
Incubate at room temperature for 20 minutes to ensure complete dissolution. If solubility issues persist, incubate overnight at 4°C.
Aliquoting and Storage:
Aliquot the reconstituted protein into small working volumes to avoid repeated freeze-thaw cycles.
Store aliquots at –20°C or –80°C for long-term storage. Avoid multiple freeze-thaw cycles, as this can reduce activity.
Preparation for Cell Culture:
Before adding to cell culture, dilute the stock solution into your culture medium to the desired final concentration (e.g., 10–100 ng/mL, depending on your assay).
If using acidic reconstitution buffers (e.g., acetic acid or HCl), ensure the final dilution into culture medium neutralizes the pH to avoid cytotoxicity.
Summary Table:
Step
Details
Centrifuge vial
20–30 sec, microcentrifuge
Solvent
10–20 mM acetic acid, or sterile water with 0.1% HSA/BSA, or 4 mM HCl
Concentration
≥100 μg/mL (0.1–1.0 mg/mL)
Mixing
Gentle swirling/tapping; no vortexing
Incubation
20 min at RT; overnight at 4°C if needed
Aliquoting
Small volumes, avoid repeated freeze-thaw
Storage
–20°C or –80°C
Cell culture use
Dilute to working concentration in medium; neutralize pH if acidic
Key technical notes:
Always consult the specific product datasheet for any unique recommendations.
Confirm protein integrity by SDS-PAGE if needed.
Use carrier protein (HSA/BSA) to minimize loss from adsorption and enhance stability.
These steps will help ensure BMP-7 retains its biological activity and is suitable for cell culture applications.
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
