Recombinant Human Sonic Hedgehog/Shh N-Terminus GMP Protein: R&D Systems

Recombinant Human BMP-4 GMP Protein, CF

  
  • Purity
    >95%, by SDS-PAGE with silver staining, under reducing conditions.
  • Endotoxin Level
    <0.01 EU per 1 μg of the protein by the LAL method.
  • Activity
    Measured by its ability to induce alkaline phosphatase production by ATDC5 mouse chondrogenic cells. Binnerts, M.E. et al. (2004) Biochem. Biophys. Res. Commun. 315(2):272. The ED50 for this effect is 2.5-15 ng/mL.
  • Source
    Mouse myeloma cell line, NS0-derived Ser293-Arg408 Manufactured and tested under current Good Manufacturing Practice (GMP) guidelines.
  • Accession #
  • N-terminal Sequence
    Analysis
    Ser293-Pro-Lys-His-His-Ser-Gln-Arg-Ala-Arg
  • Structure / Form
    Disulfide-linked homodimer
  • Predicted Molecular Mass
    13 kDa (monomer)
  • SDS-PAGE

    22-25 kDa, reducing conditions

    37-41 kDa, non-reducing conditions

314-GMP
 
Formulation Lyophilized from a 0.2 μm filtered solution in Acetonitrile and TFA.
Reconstitution Reconstitute at 50-200 μg/mL in 4 mM HCl.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • A minimum of 6 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, ≤ -20 °C under sterile conditions after reconstitution.
Data Images

GMP-grade Recombinant Human BMP-4 (Catalog # 314-GMP) induces alkaline phosphatase production in the ATDC5 mouse chondrogenic cell line. The ED50 for this effect is 2.5-15 ng/mL.

1 μg/lane of GMP-grade Recombinant Human BMP-4 (Catalog # 314-GMP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silver staining, showing bands at 19 and 21 kDa, and 35 and 38 kDa, respectively.

Background: BMP-4

BMP-4 is a TGF-beta superfamily ligand that is widely expressed from early embryogenesis through adulthood. It plays an important role in mesenchyme formation, epidermal determination, suppression of neural induction, the development of multiple organs, and tissue repair (1-5). The human BMP-4 precursor contains a 273 amino acid (aa) propeptide and a 116 aa mature protein (6). Processing of the propeptide by furin or proprotein convertase 6 enables the formation of the mature disulfide-linked homodimeric BMP-4 and facilitates its secretion. Similar intracellular processes may lead to the formation and recreation of BMP4/BMP7
disulfide-linked heterodimer (7-9). Mature human and mouse BMP-4 share 98% aa sequence identity. Human BMP-4 shares 85% aa sequence identity with human BMP-2 and less than 50% with other human BMPs. Compared to BMP-4 homodimers, BMP-4/BMP-7 heterodimers exhibit a greater potency in inducing osteogenic differentiation (9). In Xenopus, the heterodimers can also induce the formation of mesoderm, whereas BMP-4 homodimers only provide ventralizing signals for existing mesoderm (10). BMP-4 signals through tetrameric complexes composed of type I (primarily Activin RIA or BMPR-IA) and type II (primarily Activin RIIA or BMPR-II) receptors (11, 12). The bioavailability of BMP-4 is regulated by its interaction with multiple proteins and glycosaminoglycans (13-15).

 

  • References:
    1. Zhang, P. et al. (2008) Blood 111:1933.
    2. Gambaro, K. et al. (2006) Cell Death Differ. 13:1075.
    3. Simic, P. and S. Vukicevic (2005) Cytokine Growth Factor Rev. 16:299.
    4. Sadlon, T.J. et al. (2004) Stem Cells 22:457.
    5. Frank, D.B. et al. (2005) Circ. Res. 97:496.
    6. Wozney, J. et al. (1988) Science 242:1528.
    7. Cui, Y. et al. (1998) EMBO J. 17:4735.
    8. Cui, Y. et al. (2001) Genes Dev. 15:2797.
    9. Aono, A. et al. (1995) Biochem. Biophys. Res. Commun. 210:670.
    10. Nishimatsu, S. and G.H. Thomsen (1998) Mech. Dev. 74:75.
    11. Chen, D. et al. (2004) Growth Factors 22:233.
    12. Lavery, K. et al. (2008) J. Biol. Chem. April 24 epub.
    13. Rosen, V. (2006) Ann. N.Y. Acad. Sci. 1068:19.
    14. Jones, C.M. and J.C. Smith (1998) Dev. Biol. 194:12.
    15. Takada, T. et al. (2003) J. Biol. Chem. 278:43229.
  • Long Name:
    Bone Morphogenetic Protein 4
  • Entrez Gene IDs:
    652 (Human); 12159 (Mouse); 25296 (Rat); 30612 (Zebrafish)
  • Alternate Names:
    BMP-2B; BMP2B1; BMP2BMCOPS6; BMP4; BMP-4; Bone morphogenetic protein 2B; bone morphogenetic protein 4; DVR4; OFC11; ZYME

GMP Proteins
R&D Systems' GMP proteins are produced according to relevant sections of the following documents: WHO TRS, No. 822, 1992 Annex 1, Good Manufacturing Practices for Biological Products; USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and USP Chapter 92, Growth Factors and Cytokines Used in Cell Therapy Manufacturing.

R&D Systems' quality focus includes:
  • Manufactured and tested under an ISO 9001:2008 and ISO 13485:2003 certified quality system
  • Documented processes and QA control of documentation and process changes
  • Personnel training programs
  • Raw material testing and vendor qualification/monitoring
  • Fully validated equipment, processes and test methods
  • Equipment calibration schedules using a computerized calibration program
  • Facility maintenance, safety programs and pest control
  • Material review process for variances
  • Monitoring of stability over product shelf-life

R&D Systems strives to provide our customers with the analytical characteristics of each product so that customers may determine whether our products are appropriate for their research. The Certificate of Analysis provided contains the following lot specific information:

  • N-terminal amino acid analysis, SDS-PAGE analysis, and endotoxin level (as determined by LAL assay) performed on each bulk QC lot, not on individual bottlings of each QC lot
  • Post-bottling lot-specific bioassay results (compliance with an established range) and results of microbial bioburden testing (using broth culture, Sabourand's dextrose and blood agar plates with results reported at 3 days and at 7 days)

Additional testing and documentation requested by the customer can be arranged at an additional cost. Testing may include, but is not limited to, USP sterility testing, positive identity testing, testing for adventitious agents and testing for residual host cell content.

Production records and facilities are available for examination by appropriate personnel on-site at R&D Systems in Minneapolis, Minnesota USA.

R&D Systems sells its GMP grade recombinant protein products for research use or further manufacturing use in ex vivo cell therapy applications. They are not for in vivo use or for use as therapeutic or other drugs, biologic products or devices. Please read the following End User Terms prior to using this product.

END USER TERMS OF USE OF PRODUCT
The following terms are offered to you upon your acceptance of these End User Terms of Use of Product. By using this product, you indicate your acknowledgment and agreement to these End User Terms of Use of Product. If you do not agree to be bound by and comply with all of the provisions of these End User Terms of Use of Product, you should contact your supplier of the product and make arrangements to return the product.

We suggest you print and retain a copy of these End User Terms of Use of Product for your records.

The End User is aware that R&D Systems, Inc. sells its GMP products for research use only or further manufacturing and not for in vivo use, the production of therapeutics or other drugs or for biologic products or devices. The End User further agrees, as a condition of the sale of R&D Systems'' GMP products that: a) the End User will not use this GMP Product in any procedure wherein the product may be directly or indirectly administered to humans, unless the End User has obtained, or prior to their use will have obtained, an Investigational New Drug (IND) exemption from the FDA and will use the product only in accordance with the protocols of such IND and of the Institutional Review Board overseeing the proposed research, or b) the End User will use the products outside of the United States in accordance with the protocols of research approved by the Institutional Review Board or authorized ethics committee and regulatory agencies to which the End User is subject to in their territory.

R&D Systems, Inc. has the right, at its sole discretion, to modify, add or remove any terms or conditions of these End User Terms of Use without notice or liability to you. Any changes to these End User Terms of Use are effective immediately following the printing of such changes on this product insert. The most recent version of these End User Terms of Use of Product may be found at: RnDSystems.com/Legal-Information

You agree to review these End User Terms of Use of Product to ensure any subsequent use by you of R&D Systems'' GMP Products following changes to these End User Terms of Use of Product constitutes your acceptance of all such changes.

TERMS AND CONDITIONS
The following limitation applies to R&D Systems'' warranty and liability for damages: All products are warranted to meet R&D Systems'' published specifications when used under normal laboratory conditions.

R&D SYSTEMS DOES NOT MAKE ANY OTHER WARRANTY OR REPRESENTATION WHATSOEVER, WHETHER EXPRESS OR IMPLIED, WITH RESPECT TO ITS PRODUCTS. IN PARTICULAR, R&D SYSTEMS DOES NOT MAKE ANY WARRANTY OF SUITABILITY, NONINFRINGEMENT, MERCHANTABILITY OR FITNESS FOR ANY PARTICULAR PURPOSE.

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Full details of R&D Systems'' Terms and Conditions of Sale can be found online at: RnDSystems.com/Legal-Information

 
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